Pyrazole derivatives as p38 kinase inhibitors

ABSTRACT

A class of pyrazole derivatives is described for use in treating p38 kinase mediated disorders. Compounds of particular interest are defined by Formula I ##STR1## wherein Q, R 1 , R 2 , R 3  and R 4  are as described in the specification.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority from U.S. Provisional Application Ser.No. 60/047,569 filed May 22, 1997.

FIELD OF THE INVENTION

This invention relates to a novel group of pyrazole compounds,compositions and methods for treating p38 kinase mediated disorders.

BACKGROUND OF THE INVENTION

Mitogen-activated protein kinases (MAP) is a family of proline-directedserine/threonine kinases that activate their substrates by dualphosphorylation. The kinases are activated by a variety of signalsincluding nutritional and osmotic stress, UV light, growth factors,endotoxin and inflammatory cytokines. The p38 MAP kinase group is a MAPfamily of various isoforms, including p38α, p38β and p38γ, and isresponsible for phosphorylating and activating transcription factors(e.g. ATF2, CHOP and MEF2C) as well as other kinases (e.g. MAPKAP-2 andMAPKAP-3). The p38 isoforms are activated by bacteriallipopolysaccharide, physical and chemical stress and by pro-inflammatorycytokines, including tumor necrosis factor (TNF-α) and interleukin-1(IL-1). The products of the p38 phosphorylation mediate the productionof inflammatory cytokines, including TNF and IL-1, and cyclooxygenase-2.

TNF-α is a cytokine produced primarily by activated monocytes andmacrophages. Excessive or unregulated TNF production has been implicatedin mediating a number of diseases. Recent studies indicate that TNF hasa causative role in the pathogenesis of rheumatoid arthritis. Additionalstudies demonstrate that inhibition of TNF has broad application in thetreatment of inflammation, inflammatory bowel disease, multiplesclerosis and asthma.

TNF has also been implicated in viral infections, such as HIV, influenzavirus, and herpes virus including herpes simplex virus type-1 (HSV-1),herpes simplex virus type-2 (HSV-2), cytomegalovirus (CMV),varicella-zoster virus (VZV), Epstein-Barr virus, human herpesvirus-6(HHV-6), human herpesvirus-7 (HHV-7), human herpesvirus-8 (HHV-8),pseudorabies and rhinotracheitis, among others.

IL-8 is another pro-inflammatory cytokine, which is produced bymononuclear cells, fibroblasts, endothelial cells, and keratinocytes,and is associated with conditions including inflammation.

IL-1 is produced by activated monocytes and macrophages and is involvedin the inflammatory response. IL-1 plays a role in manypathophysiological responses including rheumatoid arthritis, fever andreduction of bone resorption.

TNF, IL-1 and IL-8 affect a wide variety of cells and tissues and areimportant inflammatory mediators of a wide variety of disease states andconditions. The inhibition of these cytokines by inhibition of the p38kinase is of benefit in controlling, reducing and alleviating many ofthese disease states.

Various pyrazoles have previously been described. U.S. Pat. No.4,000,281, to Beiler and Binon, describes 4,5-aryl/heteroarylsubstituted pyrazoles with antiviral activity against both RNA and DNAviruses such as myxoviruses, adenoviruses, rhinoviruses, and variousviruses of the herpes group. WO 92/19615, published Nov. 12, 1992,describes pyrazoles as novel fungicides. U.S. Pat. No. 3,984,431, toCueremy and Renault, describes derivatives of pyrazole-5-acetic acid ashaving anti-inflammatory activity. Specifically,[1-isobutyl-3,4-diphenyl-1H-pyrazol-5-yl]acetic acid is described. U.S.Pat. No. 3,245,093 to Hinsgen et al, describes a process for preparingpyrazoles. WO 83/00330, published Feb. 3, 1983, describes a new processfor the preparation of diphenyl-3,4-methyl-5-pyrazole derivatives. WO95/06036, published Mar. 2, 1995, describes a process for preparingpyrazole derivatives. U.S. Pat. No. 5,589,439, to T. Goto, et al.,describes tetrazole derivatives and their use as herbicides. EP 515041describes pyrimidyl substituted pyrazole derivatives as novelagricultural fungicides. Japanese Patent 4,145,081 describespyrazolecarboxylic acid derivatives as herbicides. Japanese Patent5,345,772 describes novel pyrazole derivatives as inhibitingacetylcholinesterase.

Pyrazoles have been described for use in the treatment of inflammation.Japanese Patent 5,017,470 describes synthesis of pyrazole derivatives asanti-inflammatory, anti-rheumatic, anti-bacterial and anti-viral drugs.EP 115640, published Dec. 30, 1983, describes 4-imidazolyl-pyrazolederivatives as inhibitors of thromboxane synthesis.3-(4-Isopropyl-1-methylcyclohex-1-yl)-4-(imidazol-1-yl)-1H-pyrazole isspecifically described. WO 97/01551, published Jan. 16, 1997, describespyrazole compounds as adenosine antagonists.4-(3-Oxo-2,3-dihydropyridazin-6-yl)-3-phenylpyrazole is specificallydescribed. U.S. Pat. No. 5,134,142, to Matsuo et al. describes1,5-diaryl pyrazoles as having anti-inflammatory activity.

U.S. Pat. No. 5,559,137 to Adams et al, describes novel pyrazoles(1,3,4,-substituted) as inhibitors of cytokines used in the treatment ofcytokine diseases. Specifically,3-(4-fluorophenyl)-1-(4-methylsulfinylphenyl)-4-(4-pyridyl)-5H-pyrazoleis described. WO 96/03385, published Feb. 8, 1996, describes3,4-substituted pyrazoles, as having anti-inflammatory activity.Specifically,4-[1-ethyl-4-(4-pyridyl)-5-trifluoromethyl-1H-pyrazol-3-yl]benzenesulfonamideis described.

The invention's pyrazolyl compounds are found to show usefulness as p38kinase inhibitors.

DESCRIPTION OF THE INVENTION

A class of substituted pyrazolyl compounds useful in treating p38mediated disorders is defined by Formula I: ##STR2## wherein R¹ isselected from hydrido, alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, aryl, heterocyclyl, cycloalkylalkylene, cycloalkenylalkylene,haloalkyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, aralkyl,aralkenyl, aralkynyl, heterocyclylalkylene, alkoxyalkyl, aryloxyalkyl,heterocyclyloxyalkyl, mercaptoalkyl, mercaptoaryl, mercaptoheterocyclyl,alkylthioalkylene, arylthioalkylene, amino, alkylamino, arylamino,aminoalkyl, aminoaryl, alkylaminoalkylene, and heterocyclylalkylene; and

Q is selected from oxy, thio, alkylene, alkenylene, alkynylene,sulfinyl, sulfonyl, ##STR3## wherein ##STR4## represents a four to eightmembered ring heterocyclylidenyl comprising one or more heteroatomsselected from oxygen, sulfur and nitrogen; and

wherein n is an integer from 1 to 7; and

R¹ is aryl optionally substituted with one or more radicalsindependently selected from halo, alkyl, alkenyl, alkynyl, aryl,heterocyclyl, alkoxy, alkenoxy, alkynoxy, aryloxy, heterocyclyloxy,aralkoxy, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl,alkylsulfonyl, arylsulfonyl, amino, alkylamino, alkenylamino,alkynylamino, arylamino, heterocyclylamino, aminoalkyl, aminocarbonyl,cyano, hydroxyl, hydroxyalkyl, alkoxycarbonyl, aryloxycarbonyl,heterocyclyloxycarbonyl, formyl, nitro, nitroalkyl, alkylcarbonylamino,arylcarbonylamino, haloalkylsulfinyl, haloalkylsulfonyl, alkylcarbonyl,arylcarbonyl, heterocyclylcarbonyl, and haloalkyl; and

R³ is heteroaryl optionally substituted with one or more radicalsindependently selected from halo, alkyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, amino, aminocarbonyl, cyano, hydroxyl,alkoxycarbonyl, formyl, aralkyl, aralkyloxy, aralkylthio, aralkylamino,aminosulfonyl, alkylamino, nitro, arylamino, alkylcarbonylamino,halosulfonyl, aminoalkyl, haloalkyl and alkylcarbonyl; and

R⁴ is selected from hydrido, alkyl, aryl, haloalkyl, heterocyclyl,cycloalkyl, alkenyl, cycloalkenyl, alkoxy, alkylthio, arylthio, carboxy,alkoxycarbonyl, carboxyalkyl, alkoxycarbonylalkylene, heterocyclylalkyl,amino, alkylamino, alkynylamino, arylamino, heterocyclylamino,heterocyclylalkylamino, heterocyclylaminoalkyl, andaminoheterocyclylamino; wherein the aryl, heterocyclyl, cycloalkyl,cycloalkenyl groups are optionally substituted with one or more radicalsindependently selected from halo, amino, alkyl, alkenyl, alkynyl,alkoxy, aryloxy, aralkoxy, haloalkyl, and alkylamino; and wherein theamino radicals of the heterocylcylalkylamino and heterocylcylaminoalkylgroup are optionally substituted with one or more alkyl; and

R⁶ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R⁷ and R⁸ are independently selected from hydrido, alkyl, alkenyl, andalkynyl, or together form a carbocyclic or heterocyclic ring havingthree to eight members; and

R⁹ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R¹⁰ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R¹¹ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R¹² is selected from hydrido and alkyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

Compounds of Formula I would be useful for, but not limited to, thetreatment of any disorder or disease state in a human, or other mammal,which is exacerbated or caused by excessive or unregulated TNF or p38kinase production by such mammal. Accordingly, the present inventionprovides a method of treating a cytokine-mediated disease whichcomprises administering an effective cytokine-interfering amount of acompound of Formula I, or a pharmaceutically acceptable salt or tautomerthereof.

Compounds of Formula I would be useful for, but not limited to, thetreatment of inflammation in a subject, and for use as antipyretics forthe treatment of fever. Compounds of the invention would be useful totreat arthritis, including but not limited to, rheumatoid arthritis,spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupuserythematosus and juvenile arthritis, osteoarthritis, gouty arthritisand other arthritic conditions. Such compounds would be useful for thetreatment of pulmonary disorders or lung inflammation, including adultrespiratory distress syndrome, pulmonary sarcoidosis, asthma, silicosis,and chronic pulmonary inflammatory disease. The compounds are alsouseful for the treatment of viral and bacterial infections, includingsepsis, septic shock, gram negative sepsis, malaria, meningitis,cachexia secondary to infection or malignancy, cachexia secondary toacquired immune deficiency syndrome (AIDS), AIDS, ARC (AIDS relatedcomplex), pneumonia, and herpesvirus. The compounds are also useful forthe treatment of bone resorption diseases, such as osteoporosis,endotoxic shock, toxic shock syndrome, reperfusion injury, autoimmunedisease including graft vs. host reaction and allograft rejections,cardiovascular diseases including atherosclerosis, thrombosis,congestive heart failure, and cardiac reperfusion injury, renalreperfusion injury, liver disease and nephritis, and myalgias due toinfection. The compounds are also useful for the treatment of influenza,multiple sclerosis, cancer, diabetes, systemic lupus erthrematosis(SLE), skin-related conditions such as psoriasis, eczema, burns,dermatitis, keloid formation, and scar tissue formation. Compounds ofthe invention also would be useful to treat gastrointestinal conditionssuch as inflammatory bowel disease, Crohn's disease, gastritis,irritable bowel syndrome and ulcerative colitis. The compounds wouldalso be useful in the treatment of ophthalmic diseases, such asretinitis, retinopathies, uveitis, ocular photophobia, and of acuteinjury to the eye tissue. Compounds of the invention also would beuseful for treatment of angiogenesis, including neoplasia; metastasis;ophthalmological conditions such as corneal graft rejection, ocularneovascularization, retinal neovascularization includingneovascularization following injury or infection, diabetic retinopathy,retrolental fibroplasia and neovascular glaucoma; ulcerative diseasessuch as gastric ulcer; pathological, but non-malignant, conditions suchas hemaginomas, including invantile hemaginomas, angiofibroma of thenasopharynx and avascular necrosis of bone; diabetic nephropathy andcardiomyopathy; and disorders of the female reproductive system such asendometriosis. The compounds of the invention may also be useful forpreventing the production of cyclooxygenase-2.

Besides being useful for human treatment, these compounds are alsouseful for veterinary treatment of companion animals, exotic animals andfarm animals, including mammals, rodents, and the like. More preferredanimals include horses, dogs, and cats.

The present compounds may also be used in co-therapies, partially orcompletely, in place of other conventional anti-inflammatories, such astogether with steroids, cyclooxygenase-2 inhibitors, NSAIDs, DMARDS,immunosuppressive agents, 5-lipoxygenase inhibitors, LTB₄ antagonistsand LTA₄ hydrolase inhibitors.

As used herein, the term "TNF mediated disorder" refers to any and alldisorders and disease states in which TNF plays a role, either bycontrol of TNF itself, or by TNF causing another monokine to bereleased, such as but not limited to IL-1, IL-6 or IL-8. A disease statein which, for instance, IL-1 is a major component, and whose productionor action, is exacerbated or secreted in response to TNF, wouldtherefore be considered a disorder mediated by TNF.

As used herein, the term "p38 mediated disorder" refers to any and alldisorders and disease states in which p38 plays a role, either bycontrol of p38 itself, or by p38 causing another factor to be released,such as but not limited to IL-1, IL-6 or IL-8. A disease state in which,for instance, IL-1 is a major component, and whose production or action,is exacerbated or secreted in response to p38, would therefore beconsidered a disorder mediated by p38.

As TNF-β has close structural homology with TNF-α (also known ascachectin), and since each induces similar biologic responses and bindsto the same cellular receptor, the synthesis of both TNF-α and TNF-β areinhibited by the compounds of the present invention and thus are hereinreferred to collectively as "TNF" unless specifically delineatedotherwise.

A preferred class of compounds consists of those compounds of Formula Iwherein

R¹ is selected from hydrido, lower alkyl, lower alkynyl, lowercycloalkylalkylene, lower haloalkyl, lower hydroxyalkyl, loweralkoxyalkyl, lower thioalkyl, lower alkylthioalkylene, amino, loweralkylamino, lower arylamino, lower alkylaminoalkylene, and lowerheterocyclylalkylene; and

Q is selected from lower alkylene, lower alkenylene, sulfinyl, sulfonyl,##STR5## wherein ##STR6## represents a four to eight membered ringheterocyclylidenyl comprising one or more heteroatoms selected fromoxygen, sulfur and nitrogen; and

wherein n is an integer from 1 to 7; and

R² is aryl optionally substituted with one or more radicalsindependently selected from halo, lower alkyl, lower alkoxy, loweraryloxy, lower aralkoxy, amino, hydroxyl, nitro, cyano, lower haloalkyl,lower alkylamino, and lower alkynylamino; and

R³ is selected from 5- to 10-membered heterocyclyl optionallysubstituted with one or more radicals independently selected from loweralkylthio, lower alkylsulfonyl, aminosulfonyl, halo, lower alkyl, loweralkylsulfinyl, cyano, lower alkoxycarbonyl, aminocarbonyl, formyl, loweraralkyl, lower aralkyloxy, lower aralkylthio, lower aralkylamino, loweralkylcarbonylamino, lower haloalkyl, hydroxyl, lower alkoxy, amino,lower alkylamino, lower aminoalkyl, phenylamino, nitro, halosulfonyl andlower alkylcarbonyl; and

R⁴ is selected from hydrido, lower alkyl, aryl, lower haloalkyl, 5-10membered heterocyclyl, lower alkylamino, lower alkynylamino,phenylamino, lower cycloalkyl, lower alkenyl, lower cycloalkenyl, loweralkoxy, lower alkylthio, carboxy, lower alkoxycarbonyl, lowercarboxyalkyl, lower alkoxycarbonylalkylene, lower heterocyclylalkyl,lower heterocylcylalkylamino, and lower heterocylcylaminoalkyl; whereinthe aryl, 5-10 membered heteroaryl, lower cycloalkyl and lowercycloalkenyl groups are optionally substituted with one or more radicalsindependently selected from halo, lower alkyl, lower alkenyl, loweralkynyl, alkoxy, phenoxy, lower aralkoxy, lower haloalkyl, and loweralkylamino; and wherein the amino radicals of the lowerheterocylcylalkylamino and lower heterocylcylaminoalkyl group areoptionally substituted with one or more lower alkyl; and

R⁶ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R⁷ and R⁸ are independently selected from hydrido, lower lower alkyl,lower alkenyl, and lower alkynyl, or together form a carbocyclic orheterocyclic ring having three to eight members; and

R⁹ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R¹⁰ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R¹¹ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R¹² is selected from hydrido and lower alkyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

A more preferred class of compounds consists of those compounds ofFormula I wherein

R¹ is selected from hydrido, lower alkyl, lower alkynyl, lowercycloalkylalkylene, lower haloalkyl, lower hydroxyalkyl, loweralkoxyalkyl, lower thioalkyl, lower alkylthioalkylene, loweralkylaminoalkylene, and lower heterocyclylalkylene; and

Q is selected from lower alkylene, lower alkenylene, ##STR7## wherein##STR8## represents a four to eight membered ring heterocyclylidenylcomprising one or more heteroatoms selected from oxygen, sulfur andnitrogen; and

wherein n is an integer from 1 to 7; and

R² is aryl optionally substituted with one or more radicalsindependently selected from halo, lower alkyl, lower alkoxy, loweraryloxy, lower aralkoxy, amino, hydroxyl, nitro, cyano, lower haloalkyl,lower alkylamino, and lower alkynylamino; and

R³ is 6-membered heteroaryl optionally substituted with one or moreradicals independently selected from halo, lower alkyl, cyano,phenethyl, benzyl, benzyloxy, benzylthio, benzylamino, phenethylamino,aminocarbonyl, lower alkylcarbonylamino, hydroxyl, amino, loweralkylamino, lower aminoalkyl, and phenylamino; and

R⁴ is selected from hydrido, lower alkyl, phenyl, lower haloalkyl, 5-10membered heterocyclyl, lower alkylamino, lower alkynylamino,phenylamino, lower cycloalkyl, lower alkenyl, lower cycloalkenyl, loweralkoxy, lower heterocyclylaminoalkyl, and lower heterocyclylalkylamino;wherein the phenyl, 5-10 membered heteroaryl, lower cycloalkyl and lowercycloalkenyl groups are optionally substituted with one or more radicalsindependently selected from halo, lower alkyl, lower alkenyl, loweralkynyl, lower alkoxy, phenoxy, lower aralkoxy, lower haloalkyl, amino,hydroxyl, cyano and lower alkylamino; and wherein the amino radicals oflower heterocyclyl alkylamino and lower heterocyclylaminoalkyl areoptionally substituted with one or more lower alkyl; and

R⁶ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R⁷ and R⁸ are independently selected from hydrido, lower lower alkyl,lower alkenyl, and lower alkynyl, or together form a carbocyclic orheterocyclic ring having three to eight members; and

R⁹ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R¹⁰ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R¹¹ is selected from hydrido, lower alkyl, lower alkenyl, and loweralkynyl; and

R¹² is selected from hydrido and lower alkyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

A class of compounds of particular interest consists of those compoundsof Formula I wherein

R¹ is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, isobutyl, ethynyl, propargyl, fluoromethyl, difluoromethyl,trifluoromethyl, chloromethyl, dichloromethyl, trichloroethyl,pentafluoroethyl, heptafluoropropyl, difluorochloromethyl,dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl,dichloropropyl, morpholinomethyl, pyrrolidinylmethyl, piperazinylmethyl,piperidinylmethyl, pyridylmethyl, thienylmethyl, methoxymethyl,ethoxymethyl, methylaminomethyl, cyclohexylmethyl, hydroxymethyl,hydroxylethyl, thiomethyl, and methylthiomethyl; and

Q is selected from methylene, ethylene, propylene, ethenylene,propenylenyl ##STR9## wherein ##STR10## represents a four to eightmembered ring heterocyclylidenyl comprising one or more heteroatomsselected from oxygen, sulfur and nitrogen; and

wherein n is an integer from 1 to 7; and

R² is phenyl optionally substituted with one or more radicalsindependently selected from fluoro, chloro, bromo, methyl, ethyl,isopropyl, tert-butyl, isobutyl, methoxy, ethoxy, phenoxy, benzyloxy,trifluoromethyl, is fluoromethyl, difluoromethyl, amino, cyano, nitro,dimethylamino, ethynylamino, propargylamino, and hydroxyl; and

R³ is selected from pyridyl, pyridium, and pyrimidyl; wherein R³ isoptionally substituted with one or more radicals independently selectedfrom fluoro, chloro, bromo, methyl, ethyl, isopropyl, cyano,aminocarbonyl, methylcarbonylamino, hydroxy, benzyl, phenethyl,methylamino, ethylamino, dimethylamino, diethylamino, aminomethyl,aminoethyl, N-methyl-N-phenylamino, phenylamino, diphenylamino,benzylamino, phenethylamino, and amino; and

R⁴ is selected from hydrido, methyl, ethyl, propyl, isopropyl, butyl,tert-butyl, isobutyl, phenyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cyclopropylenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl,cyclohexadienyl, pyridyl, thienyl, isothiazolyl, isoxazolyl, thiazolyl,oxazolyl, pyrimidinyl, quinolyl, isoquinolinyl, imidazolyl,benzimidazolyl, furyl, benzofuryl, methoxy, ethoxy, trifluoromethyl,fluoromethyl, methylamino, ethynylamino, propargylamino, piperidinyl,piperazinyl, piperadinylmethyl, piperadinylmethylamino,piperadinylaminomethyl, piperazinylmethyl, piperazinylmethylamino,piperazinylaminomethyl; wherein the phenyl piperadinyl and piperazinylgroups are optionally substituted with one or more radicalsindependently selected from fluoro, chloro, bromo, methyl, ethyl,isopropyl, tert-butyl, isobutyl, propargyl, methoxy, ethoxy, phenoxy,benzyloxy, trifluoromethyl, fluoromethyl, difluoromethyl, amino,hydroxyl, cyano and dimethylamino; and wherein the amino radicals ofpiperadinylmethylamino, piperadinylaminomethyl, piperazinylmethylamino,and piperazinylaminomethyl are optionally substituted with one or moremethyl; and

R⁶ is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, and isobutyl; and

R⁷ and R⁸ are independently selected from hydrido, methyl, ethyl,propyl, isopropyl, tert-butyl, and isobutyl, or together form acarbocyclic or heterocyclic ring having three to eight members; and

R⁹ is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, and isobutyl; and

R¹⁰ is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, and isobutyl; and

R¹¹ is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, and isobutyl; and

R¹² is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, and isobutyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

A class of compounds of specific interest consists of those compounds ofFormula I wherein

R¹ is hydrido or methyl; and

Q is selected from methylene, ethylene, ethenylene, ##STR11## wherein##STR12## represents a four to eight membered ring heterocyclylidenylcomprising one or more heteroatoms selected from oxygen, sulfur andnitrogen; and

wherein n is an integer from 1 to 3; and

R² is phenyl optionally substituted with one or more radicalsindependently selected from fluoro, chloro, bromo, methyl, ethyl,isopropyl, tert-butyl, isobutyl, methoxy, ethoxy, phenoxy, benzyloxy,trifluoromethyl, fluoromethyl, difluoromethyl, amino, cyano, nitro,dimethylamino, and hydroxyl; and

R³ is pyridyl optionally substituted with one or more radicalsindependently selected from fluoro, chloro, bromo, methyl, cyano,benzyl, phenethyl, aminocarbonyl, hydroxyl, dimethylamino, benzylamino,phenethylamino, aminomethyl and amino; and

R⁴ is selected from hydrido, methyl, ethyl, propyl, propargylamino, andphenyl optionally substituted with one or more radicals independentlyselected from fluoro, chloro, bromo, methyl, ethyl, isopropyl, methoxy,ethoxy, phenoxy, benzyloxy, trifluoromethyl, dimethylamino, ethynylaminoand propargylamino; and

R⁶ is selected from hydrido and methyl; and

R⁷ and R⁸ are independently selected from hydrido and methyl; and

R⁹ is selected from hydrido and methyl; and

R¹⁰ is selected from hydrido and methyl; and

R¹¹ is selected from hydrido and methyl; and

R¹² is selected from hydrido and methyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

Within Formula I there is a subclass of compounds of high interestrepresented by Formula II: ##STR13## wherein R¹ is selected from hydridoand lower alkyl; and

Q is selected from lower alkylene, lower alkenylene, ##STR14## wherein##STR15## represents a four to eight membered ring heterocyclylidenylcomprising one or more heteroatoms selected from oxygen, sulfur andnitrogen; and

wherein n is an integer from 1 to 4; and

R² is phenyl optionally substituted with one or more radicalsindependently selected from halo and lower alkyl; and

R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, and R¹² are independently selectedfrom hydrido and lower alkyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

A preferred class of compounds consists of those compounds of Formula IIwherein

R¹ is selected from hydrido and methyl; and

wherein Q is selected from methylene, ethylene, ##STR16## ethenylene, R²is phenyl optionally substituted with one or more radicals independentlyselected from fluoro, chloro and bromo; and

R⁴ is selected from hydrido, methyl and ethyl; and

R⁵ is selected from hydrido and methyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

A family of specific compounds of particular interest within Formula Iconsists of compounds, tautomers and pharmaceutically-acceptable saltsthereof as follows:

4-[3-methyl-5-(2-phenylethenyl)-1H-pyrazol-4-yl]pyridine;

4-[3-methyl-5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine;

4-[3-methyl-5-[2-(3-fluorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

4-[3-[2-(3-fluorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

4-[1-methyl-3-[2-(3-fluorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

4-[3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

4-[1-methyl-3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[l-methyl-3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

4-[3-[2-(3-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-[2-(3,4-dichlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[l-methyl-3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;

4-[3-methyl-5-[2-(3-florophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

4-[3-[2-(3-fluorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

4-[1-methyl-3-[2-(3-fluorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

4-[3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

4-[1-methyl-3-[2-(4-chiorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

4-[3-[2-(3-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-[2-(3,4-dichlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;

4-[3-methyl-5-(3-fluorobenzyl)-1H-pyrazol-4-yl]pyridine;

4-[3-(3-fluorobenzyl)-1H-pyrazol-4-yl]pyridine;

4-[1-methyl-3-(3-fluorobenzyl)-1H-pyrazol-4-yl]pyridine;

4-[3-benzyl-1H-pyrazol-4-yl]pyridine;

4-[3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;

4-[1-methyl-3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;

4-[3-(3-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-(3,4-dichlorobenzyl)-1H-pyrazol-4-yl]pyridine;

3-methyl-4-[1-methyl-3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;

(3-fluorophenyl)[3-methyl-4-(4-pyridinyl)-1H-pyrazol-5-yl]methanone;

(3-fluorophenyl)[4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(3-fluorophenyl)[1-methyl-4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;

phenyl[1-methyl-4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(4-chlorophenyl)[4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(4-chlorophenyl)[1-methyl-4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(4-chlorophenyl)[1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(3-chlorophenyl)[4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(3,4-dichlorophenyl)[1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazol-3-yl]methanone;

(4-chlorophenyl)[1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazol-3-yl]methanone;

α-(3-fluorophenyl)-3-methyl-4-(4-pyridinyl)-1H-pyrazole-5-methanol;

α-(3-fluorophenyl)-4-(4-pyridinyl)-1H-pyrazole-3-methanol;

α-(3-fluorophenyl)-1-methyl-4-(4-pyridinyl)-1H-pyrazole-3-methanol;

α-phenyl-1-methyl-4-(4-pyridinyl)-1H-pyrazole-3-methanol;

α-(4-chlorophenyl)-4-(4-pyridinyl)-1H-pyrazole-3-methanol;

α-(4-chlorophenyl)-1-methyl-4-(4-pyridinyl)-1H-pyrazole-3-methanol;

α-(4-chlorophenyl)-1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazole-3-methanol;

α-(3-chlorophenyl)-4-(4-pyridinyl)-1H-pyrazole-3-methanol;

α-(3,4-dichlorophenyl)-1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazole-3-methanol;

α-(4-chlorophenyl)-1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazole-3-methanol;and

4-[5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine.

The term "hydrido" denotes a single hydrogen atom (H). This hydridoradical may be attached, for example, to an oxygen atom to form ahydroxyl radical or two hydrido radicals may be attached to a carbonatom to form a methylene (--CH₂ --) radical. Where used, either alone orwithin other terms such as "haloalkyl", "alkylsulfonyl", "alkoxyalkyl"and "hydroxyalkyl", "mercaptoalkyl", the term "alkyl" embraces linear orbranched radicals having one to about twenty carbon atoms or,preferably, one to about twelve carbon atoms. More preferred alkylradicals are "lower alkyl" radicals having one to about ten carbonatoms. Most preferred are lower alkyl radicals having one to about sixcarbon atoms. Examples of such radicals include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,hexyl and the like. The term "alkylene" embraces bridging alkylradicals. The term "alkenyl" embraces linear or branched radicals havingat least one carbon-carbon double bond of two to about twenty carbonatoms or, preferably, two to about twelve carbon atoms. More preferredalkenyl radicals are "lower alkenyl" radicals having two to about sixcarbon atoms. Examples of alkenyl radicals include ethenyl, 1-propenyl,allyl, 2-propenyl, butenyl and 4-methylbutenyl. The terms "alkenyl" and"lower alkenyl", embrace radicals having "cis" and "trans" orientations,or alternatively, "E" and "Z" orientations. The term "alkenylene"describes bridging alkenyl radicals. The term "alkynyl" embraces linearor branched radicals having at least one carbon--carbon triple bond oftwo to about twenty carbon atoms or, preferably, two to about twelvecarbon atoms. More preferred alkynyl radicals are "lower alkynyl"radicals having two to about six carbon atoms. Examples of alkynylradicals include ethynyl, propynyl and propargyl. The term "alkynylene"describes bridging alkynyl radicals. The term "cycloalkyl" embracessaturated carbocyclic radicals having three to about twelve carbonatoms. More preferred cycloalkyl radicals are "lower cycloalkyl"radicals having three to about eight carbon atoms. Examples of suchradicals include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.The term "cycloalkylalkylene" embraces alkyl radicals substituted with acycloalkyl radical. More preferred cycloalkylalkylene radicals are"lower cycloalkylalkylene" which embrace lower alkyl radicalssubstituted with a lower cycloalkyl radical as defined above. Examplesof such radicals include cyclopropylmethylene, cyclobutylmethylene,cyclopentylmethylene and cyclohexylmethylene. The term "cycloalkenyl"embraces partially unsaturated carbocyclic radicals having three totwelve carbon atoms. When a cycloalkenyl radical embraces partiallyunsaturated carbocyclic radicals which contain two double bonds but notnecessary conjugated, it can be called "cycloalkyldienyl". Morepreferred cycloalkenyl radicals are "lower cycloalkenyl" radicals havingfour to about eight carbon atoms. Examples of such radicals includecyclobutenyl, cyclopentenyl and cyclohexenyl. The term "halo" meanshalogens such as fluorine, chlorine, bromine or iodine. The term"haloalkyl" embraces radicals wherein any one or more of the alkylcarbon atoms is substituted with halo as defined above. Specificallyembraced are monohaloalkyl, dihaloalkyl and polyhaloalkyl radicals. Amonohaloalkyl radical, for one example, may have either an iodo, bromo,chloro or fluoro atom within the radical. Dihalo and polyhaloalkylradicals may have two or more of the same halo atoms or a combination ofdifferent halo radicals. "Lower haloalkyl" embraces radicals having oneto six carbon atoms. Examples of haloalkyl radicals includefluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl,dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl,difluorochloromethyl, dichlorofluoromethyl, difluoroethyl,difluoropropyl, dichloroethyl and dichloropropyl. The term"hydroxyalkyl" embraces linear or branched alkyl radicals having one toabout ten carbon atoms, any one of which may be substituted with one ormore hydroxyl radicals. More preferred hydroxyalkyl radicals are "lowerhydroxyalkyl" radicals having one to six carbon atoms and one or morehydroxyl radicals. Examples of such radicals include hydroxymethyl,hydroxyethyl, hydroxypropyl, hydroxybutyl and hydroxyhexyl. The terms"alkoxy" and "alkyloxy" embrace linear or branched oxy-containingradicals each having alkyl portions of one to about ten carbon atoms.More preferred alkoxy radicals are "lower alkoxy" radicals having one tosix carbon atoms. Examples of such radicals include methoxy, ethoxy,propoxy, butoxy and tert-butoxy. The term "alkoxyalkyl" embraces alkylradicals having one or more alkoxy radicals attached to the alkylradical to form, for example, monoalkoxyalkyl and dialkoxyalkylradicals. The "alkoxy" radicals may be further substituted with one ormore halo atoms, such as fluoro, chloro or bromo, to provide"haloalkoxy" radicals.

The term "aryl", alone or in combination, means a carbocyclic aromaticsystem containing one, two or three rings wherein such rings may beattached together in a pendent manner or may be fused. More preferredaryl are 6-12 membered aryl. Examples of such radicals include, but notlimited to, phenyl, naphthyl, tetrahydronaphthyl, indane and biphenyl.Aryl moieties may also be substituted at a substitutable position withone or more substituents selected independently from halo, alkyl,alkenyl, alkynyl, aryl, heterocyclyl, alkoxy, alkenoxy, alkynoxy,aryloxy, heterocyclyloxy, aralkoxy, alkylthio, arylthio, alkylsulfinyl,arylsulfinyl, alkylsulfonyl, arylsulfonyl, amino, alkylamino,alkenylamino, alkynylamino, arylamino, heterocyclylamino, aminoalkyl,aminocarbonyl, cyano, hydroxyl, hydroxyalkyl, alkoxycarbonyl,aryloxycarbonyl, heterocyclyloxycarbonyl, formyl, nitro, nitroalkyl,alkylcarbonylamino, arylcarbonylamino, haloalkylsulfinyl,haloalkylsulfonyl, alkylcarbonyl, arylcarbonyl, heterocyclylcarbonyl,haloalkyl, alkoxyalkyl, alkylaminoalkyl, carboxyalkyl,alkoxycarbonylalkyl, aminocarbonylalkyl, acyl, carboxy, andaralkoxycarbonyl.

The term "heterocyclyl" embraces saturated, partially unsaturated andunsaturated heteroatom-containing ring-shaped radicals, which can alsobe called "heterocyclyl", "heterocycloalkenyl" and "heteroaryl"correspondingly, where the heteroatoms may be selected from nitrogen,sulfur and oxygen. Examples of saturated heterocyclyl radicals includesaturated 3 to 6-membered heteromonocyclic group containing 1 to 4nitrogen atoms (e.g. pyrrolidinyl, imidazolidinyl, piperidino,piperazinyl, etc.); saturated 3 to 6-membered heteromonocyclic groupcontaining 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms (e.g.morpholinyl, etc.); saturated 3 to 6-membered heteromonocyclic groupcontaining 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms (e.g.,thiazolidinyl, etc.). Examples of partially unsaturated heterocyclylradicals include dihydrothiophene, dihydropyran, dihydrofuran anddihydrothiazole. Heterocyclyl radicals may include a pentavalentnitrogen, such as in tetrazolium and pyridinium radicals. The term"heteroaryl" embraces unsaturated heterocyclyl radicals. Examples ofheteroaryl radicals include unsaturated 3 to 6 membered heteromonocyclicgroup containing 1 to 4 nitrogen atoms, for example, pyrrolyl,pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidyl, pyrazinyl,pyridazinyl, triazolyl (e.g., 4H-1,2,4-triazolyl, 1H-1,2,3-triazolyl,2H-1,2,3-triazolyl, etc.) tetrazolyl (e.g. 1H-tetrazolyl, 2H-tetrazolyl,etc.), etc.; unsaturated condensed heterocyclyl group containing 1 to 5nitrogen atoms, for example, indolyl, isoindolyl, indolizinyl,benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl,tetrazolopyridazinyl (e.g., tetrazolo[1,5-b]pyridazinyl, etc.), etc.;unsaturated 3 to 6-membered heteromonocyclic group containing an oxygenatom, for example, pyranyl, furyl, etc.; unsaturated 3 to 6-memberedheteromonocyclic group containing a sulfur atom, for example, thienyl,etc.; unsaturated 3- to 6-membered heteromonocyclic group containing 1to 2 oxygen atoms and 1 to 3 nitrogen atoms, for example, oxazolyl,isoxazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl,1,2,5-oxadiazolyl, etc.) etc.; unsaturated condensed heterocyclyl groupcontaining 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms (e.g.benzoxazolyl, benzoxadiazolyl, etc.); unsaturated 3 to 6-memberedheteromonocyclic group containing 1 to 2 sulfur atoms and 1 to 3nitrogen atoms, for example, thiazolyl, thiadiazolyl (e.g.,1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.) etc.;unsaturated condensed heterocyclyl group containing 1 to 2 sulfur atomsand 1 to 3 nitrogen atoms (e.g., benzothiazolyl, benzothiadiazolyl,etc.) and the like. The terms "heteroaryl and heterocyclyl" also embraceradicals where heterocyclyl radicals are fused with aryl radicals.Examples of such fused bicyclic radicals include benzofuran,benzothiophene, and the like. Said heterocyclyl group may have 1 to 3substituents such as alkyl, hydroxyl, halo, alkoxy, oxo, amino andalkylamino. The term "heterocyclylalkylene" embraces saturated,partially unsaturated and unsaturated heterocyclyl-substituted alkylradicals. More preferred heterocyclylalkylene radicals are "lowerheterocyclylalkylene" radicals having one to six carbon atoms and aheterocyclyl radical. Examples of such radicals includepyrrolidinylmethyl, pyridylmethyl, quinolylmethyl, thienylmethyl,furylethyl, and quinolylethyl. The heteroaryl group in saidheteroaralkyl may be additionally substituted with halo, alkyl, alkoxy,haloalkyl and haloalkoxy.

The term "alkylthio" embraces radicals containing a linear or branchedalkyl radical, of one to about ten carbon atoms attached to a divalentsulfur atom. More preferred alkylthio radicals are "lower alkylthio"radicals having alkyl radicals of one to six carbon atoms. Examples ofsuch lower alkylthio radicals are methylthio, ethylthio, propylthio,butylthio and hexylthio. The term "alkylthioalkylene" embraces radicalscontaining an alkylthio radical attached through the divalent sulfuratom to an alkyl radical of one to about ten carbon atoms. Morepreferred alkylthioalkylene radicals are "lower alkylthioalkylene"radicals having alkyl radicals of one to six carbon atoms. Examples ofsuch lower alkylthioalkylene radicals include methylthiomethyl. The term"alkylsulfinyl" embraces radicals containing a linear or branched alkylradical, of one to about ten carbon atoms, attached to a divalent--S(═O)-- radical. More preferred alkylsulfinyl radicals are "loweralkylsulfinyl" radicals having alkyl radicals of one to six carbonatoms. Examples of such lower alkylsulfinyl radicals includemethylsulfinyl, ethylsulfinyl, butylsulfinyl and hexylsulfinyl. The term"sulfonyl", whether used alone or linked to other terms, such as"alkylsulfonyl", or "halosulfonyl," denotes the divalent radical, --SO₂--. "Alkylsulfonyl" embraces alkyl radicals attached to a sulfonylradical, where alkyl is defined as above. More preferred alkylsulfonylradicals are "lower alkylsulfonyl" radicals having one to six carbonatoms. Examples of such lower alkylsulfonyl radicals includemethylsulfonyl, ethylsulfonyl and propylsulfonyl. The "alkylsulfonyl"radicals may be further substituted with one or more halo atoms, such asfluoro, chloro or bromo, to provide haloalkylsulfonyl radicals. The term"halosulfonyl" embraces halo radicals attached to a sulfonyl radical.Examples of such halosulfonyl radicals include chlorosulfonyl, andbromosulfonyl. The terms "sulfamyl", "aminosulfonyl" and "sulfonamidyl"denote NH₂ O₂ S--.

The term "carbonyl", whether used alone or with other terms, such as"alkoxycarbonyl", denotes ##STR17## The terms "carboxy" or "carboxyl",whether used alone or with other terms, such as "carboxyalkyl", denotes--CO₂ H. The term "carboxyalkyl" embraces alkyl radicals substitutedwith a carboxy radical. More preferred are "lower carboxyalkyl" radicalswhich embrace carboxy-substituted lower alkyl radicals as defined above.Examples of such lower carboxyalkyl radicals include carboxymethyl,carboxyethyl and carboxypropyl. The term "alkoxycarbonyl" means aradical containing an alkoxy radical, as defined above, attached via anoxygen atom to a carbonyl radical. More preferred are "loweralkoxycarbonyl" radicals with alkyl portions having one to six carbons.Examples of such lower alkoxycarbonyl (ester) radicals includemethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl andhexyloxycarbonyl. The term "alkoxycarbonylalkylene" embraces alkyleneradicals substituted with an alkoxycarbonyl radical as defined above.More preferred are "lower alkoxycarbonylalkylene" radicals with alkyleneportions having one to six carbons. Examples of such loweralkoxycarbonylalkylene radicals include substituted or unsubstitutedmethoxycarbonylmethyl, ethoxycarbonylmethyl, methoxycarbonylethyl andethoxycarbonylethyl. The term "alkylcarbonyl", includes radicals havingalkyl radicals, as defined herein, attached to a carbonyl radical.Examples of such radicals include substituted or unsubstitutedmethylcarbonyl, ethylcarbonyl, propylcarbonyl, butylcarbonyl, andpentylcarbonyl. The term "aralkyl" embraces aryl-substituted alkylradicals. Preferred are "lower aralkyl" radicals having branched orunbranched lower alkyl portions containing one to six carbon atoms.Examples include benzyl, diphenylmethyl, triphenylmethyl, phenylethyl,and diphenylethyl. The aryl in said aralkyl may be additionallysubstituted with halo, alkyl, alkoxy, haloalkyl and haloalkoxy. Theterms benzyl and phenylmethyl are interchangeable. The term "aryloxy"embraces aryl radicals attached through an oxygen atom to otherradicals. The term "aralkoxy" embraces aralkyl radicals attached throughan oxygen atom to other radicals.

The term "aminoalkyl" embraces alkyl radicals substituted with aminoradicals. More preferred are "lower aminoalkyl" radicals. Examples ofsuch radicals include aminomethyl, aminoethyl, and the like. The term"alkylamino" denotes amino groups which are substituted with one or twoalkyl radicals. Preferred are "lower alkylamino" radicals having alkylportions having one to six carbon atoms. Suitable lower alkylamino maybe monosubstituted N-alkylamino or disubstituted N,N-alkylamino, such asN-methylamino, N-ethylamino, N,N-dimethylamino, N,N-diethylamino or thelike. The term "arylamino" denotes amino groups which are substitutedwith one or two aryl radicals, such as N-phenylamino. The "arylaminol"radicals may be further substituted on the aryl ring portion of theradical. The term "aminocarbonyl" denotes an amide group of the formula--C(═O)NH₂. The term "alkylaminocarbonyl" denotes an aminocarbonyl groupwhich has been substituted with one or two alkyl radicals on the aminonitrogen atom. Preferred are "N-alkylaminocarbonyl" and"N,N-dialkylaminocarbonyl" radicals. More preferred are "lowerN-alkylaminocarbonyl" and "lower N,N-dialkylaminocarbonyl" radicals withlower alkyl portions as defined above. The term "alkylcarbonylamino"embraces amino groups which are substituted with an alkylcarbonylradical. More preferred alkylcarbonylamino radicals are "loweralkylcarbonylamino" having lower alkylcarbonyl radicals as defined aboveattached to amino radicals. The term "alkylaminoalkylene" embracesradicals having one or more alkyl radicals attached to an aminoalkylradical.

The additional terms used to describe the substituents of the pyrazolering and not specifically defined herein are defined in a similar mannerto that illustrated in the above definitions. As above, more preferredsubstituents are those containing "lower" radicals. Unless otherwisedefined to contrary, the term "lower" as used in this application meansthat each alkyl radical of a pyrazole ring substituent comprising one ormore alkyl radicals has one to about six carbon atoms; each alkenylradical of a pyrazole ring substituent comprising one or more alkenylradicals has two to about six carbon atoms; each alkynyl radical of apyrazole ring substituent comprising one or more alkynyl radicals hastwo to about six carbon atoms; each cycloalkyl or cycloalkenyl radicalof a pyrazole ring substituent comprising one or more cycloalkyl and/orcycloalkenyl radicals is a 3 to 8 membered ring cycloalkyl orcycloalkenyl radical, respectively; each aryl radical of a pyrazole ringsubstituent comprising one or more aryl radicals is a monocyclic arylradical; and each heterocyclyl radical of a pyrazole ring substituentcomprising one or more heterocyclyl radicals is a 4-8 membered ringheterocyclyl.

The present invention comprises the tautomeric forms of compounds ofFormulas I-VIII. As illustrated below, the pyrazoles of Formula I' andI" are magnetically and structurally equivalent because of theprototropic tautomeric nature of the hydrogen: ##STR18##

The present invention also comprises compounds of Formula I-VIII havingone or more asymmetric carbons. It is known to those skilled in the artthat those pyrazoles of the present invention having asymmetric carbonatoms may exist in diastereomeric, racemic, or optically active forms.All of these forms are contemplated within the scope of this invention.More specifically, the present invention includes enantiomers,diastereomers, racemic mixtures, and other mixtures thereof.

The present invention comprises a pharmaceutical composition for thetreatment of a TNF mediated disorder, a p38 kinase mediated disorder,inflammation and/or arthritis, comprising a therapeutically-effectiveamount of a compound of Formula I-VIII, or a therapeutically-acceptablesalt or tautomer thereof, in association with at least onepharmaceutically-acceptable carrier, adjuvant or diluent.

The present invention also comprises a therapeutic method of treating aTNF mediated disorder, a p38 kinase mediated disorder, inflammationand/or arthritis in a subject, the method comprising treating a subjecthaving or susceptible to such disorder or condition with atherapeutically-effective amount of a compound of Formula I, or atherapeutically-acceptable salt or tautomer thereof, wherein

R¹ is selected from hydrido, alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, aryl, heterocyclyl, cycloalkylalkylene, cycloalkenylalkylene,haloalkyl, hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, aralkyl,aralkenyl, aralkynyl, heterocyclylalkylene, alkoxyalkyl, aryloxyalkyl,heterocyclyloxyalkyl, mercaptoalkyl, mercaptoaryl, mercaptoheterocyclyl,alkylthioalkylene, arylthioalkylene, amino, alkylamino, arylamino,aminoalkyl, aminoaryl, alkylaminoalkylene, and heterocyclylalkylene; and

Q is selected from oxy, thio, alkylene, alkenylene, alkynylene,sulfinyl, sulfonyl, ##STR19## wherein ##STR20## represents a four toeight membered ring heterocyclylidenyl comprising one or moreheteroatoms selected from oxygen, sulfur and nitrogen; and

wherein n is an integer from 1 to 7; and

R² is aryl optionally substituted with one or more radicalsindependently selected from halo, alkyl, alkenyl, alkynyl, aryl,heterocyclyl, alkoxy, alkenoxy, alkynoxy, aryloxy, heterocyclyloxy,aralkoxy, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl,alkylsulfonyl, arylsulfonyl, amino, alkylamino, alkenylamino,alkynylamino, arylamino, heterocyclylamino, aminoalkyl, aminocarbonyl,cyano, hydroxyl, hydroxyalkyl, alkoxycarbonyl, aryloxycarbonyl,heterocyclyloxycarbonyl, formyl, nitro, nitroalkyl, alkylcarbonylamino,arylcarbonylamino, haloalkylsulfinyl, haloalkylsulfonyl, alkylcarbonyl,arylcarbonyl, heterocyclylcarbonyl, and haloalkyl; and

R³ is heteroaryl optionally substituted with one or more radicalsindependently selected from halo, alkyl, alkoxy, alkylthio,alkylsulfinyl, alkylsulfonyl, amino, aminocarbonyl, cyano, hydroxyl,alkoxycarbonyl, formyl, aralkyl, aralkyloxy, aralkylthio, aralkylamino,aminosulfonyl, alkylamino, nitro, arylamino, alkylcarbonylamino,halosulfonyl, aminoalkyl, haloalkyl and alkylcarbonyl; and

R⁴ is selected from hydrido, alkyl, aryl, haloalkyl, heterocyclyl,cycloalkyl, alkenyl, cycloalkenyl, alkoxy, alkylthio, arylthio, carboxy,alkoxycarbonyl, carboxyalkyl, alkoxycarbonylalkylene, heterocyclylalkyl,amino, alkylamino, alkynylamino, arylamino, heterocyclylamino,heterocyclylalkylamino, heterocyclylaminoalkyl, andaminoheterocyclylamino; wherein the aryl, heterocyclyl, cycloalkyl,cycloalkenyl groups are optionally substituted with one or more radicalsindependently selected from halo, amino, alkyl, alkenyl, alkynyl,alkoxy, aryloxy, aralkoxy, haloalkyl, and alkylamino; and wherein theamino radicals of the heterocylcylalkylamino and heterocylcylaminoalkylgroup are optionally substituted with one or more alkyl; and

R⁶ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R⁷ and R⁸ are independently selected from hydrido, alkyl, alkenyl, andalkynyl, or together form a carbocyclic or heterocyclic ring havingthree to eight members; and

R⁹ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R¹⁰ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R¹¹ is selected from hydrido, alkyl, alkenyl, and alkynyl; and

R¹² is selected from hydrido and alkyl; or

a pharmaceutically-acceptable salt or tautomer thereof.

Also included in the family of compounds of Formula I are thepharmaceutically-acceptable salts thereof. The term"pharmaceutically-acceptable salts" embraces salts commonly used to formalkali metal salts and to form addition salts of free acids or freebases. The nature of the salt is not critical, provided that it ispharmaceutically-acceptable. Suitable pharmaceutically-acceptable acidaddition salts of compounds of Formula I may be prepared from aninorganic acid or from an organic acid. Examples of such inorganic acidsare hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuricand phosphoric acid. Appropriate organic acids may be selected fromaliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclyl,carboxylic and sulfonic classes of organic acids, example of which areformic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic,tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic,aspartic, glutamic, benzoic, anthranilic, mesylic, stearic, salicylic,p-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic,toluenesulfonic, 2-hydroxyethanesulfonic, sulfanilic,cyclohexylaminosulfonic, algenic, β-hydroxybutyric, galactaric andgalacturonic acid. Suitable pharmaceutically-acceptable base additionsalts of compounds of Formula I include metallic salts and organicsalts. More preferred metallic salts include, but are not limited toappropriate alkali metal (group Ia) salts, alkaline earth metal (groupIIa) salts and other physiological acceptable metals. Such salts can bemade from aluminum, calcium, lithium, magnesium, potassium, sodium andzinc. Preferred organic salts can be made from tertiary amines andquaternary ammonium salts, including in part, tromethamine,diethylamine, N,N'-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, meglumine (N-methylglucamine) andprocaine. All of these salts may be prepared by conventional means fromthe corresponding compound of Formulas I-VIII by reacting, for example,the appropriate acid or base with the compound of Formulas I-VIII.

General Synthetic Procedures

The compounds of the invention can be prepared according to thefollowing procedures of Schemes I-X wherein the R¹ -R¹¹ and Qsubstituents are as previously defined for compounds of Formula I or IIexcept where noted. ##STR21## General Synthetic Scheme I shows thepreparation of the pyrazoles of the present invention where R¹ ishydrido and where Q is a saturated alkylene bridging radical 5 or anunsaturated alkenylene bridge 4. A propenyl derivative 1 is condensedwith heterocyclic ketones and aldehydes 2 by the Knoevenaglecondensation to give the pictured dienone 3. The dienone 3 is condensed,such as with tosyl hydrazide in boiling acetic acid, to yield thecorresponding pyrazole derivative 4 of the present invention (where R¹is hydrido, Q is alkenylene). Other compounds of the invention 5 (whereQ is alkylene) can be prepared from pyrazole 4 via hydrogenation of thedouble bond, such as with hydrogen gas and palladium on carbon in asuitable solvent, such as methanol. ##STR22##

General Synthetic Scheme II shows the preparation of a subset of thepyrazoles of the present invention (where R² is optionally substitutedphenyl, R³ is optionally substituted pyridyl, and R⁴ is hydrido) where Qis a saturated alkylene bridging radical 11 or an unsaturated alkenylenebridge 10. A cinnamic ester 6 is alkylated with the anion of 4-picoline7, such as by treatment with lithium hexamethyldisilazide (or othersuitable base, such as sodium hydride) to the pictured enone 8. Reactionof the enone 8 with dimethylformamide dimethyl acetal gives the picturedvinylamine 9, which upon reaction with a suitable hydrazine yields thepyrazole 10 having an unsaturated alkenylene bridging radical. Theunsaturated pyrazole 10 can be reduced to the corresponding saturatedpyrazole 11, such as with hydrogen gas in the presence of palladium oncarbon. ##STR23##

General Synthetic Scheme III shows the preparation of a subset of thepyrazoles of the present invention (where R¹ is hydrido, R² isoptionally substituted phenyl, R³ is optionally substituted pyridyl, R⁴is hydrido) where Q is a carbonyl radical 15, a hydroxy-substitutedalkylene bridge 16, or a saturated methylene bridging radical 17.Suitable acetophenone derivatives 12 are condensed with optionallysubstituted pyridines 13 in the presence of base to form enones 14.Cyclization of enone 14, such as with trimethylsilyldiazomethane,according to the published procedure of Aoyama, et al. (Chem. Pharm.Bull., 37, 253-256 (1989)) gives the pictured pyrazole-ketone 15. Othercompounds of the invention are produced from the ketone 15, such as byreduction with sodium borohydride in methanol to form the alcohols 16.The alcohols 16 may be hydrogenolyzed by the action of hydrogen gas andpalladium on carbon to form the pyrazoles 17 (where Q is methylene).##STR24##

General Synthetic Scheme IV shows the preparation of the pyrazoles ofthe present invention where R¹ is alkyl or substituted alkyl. Thepyrazole derivatives 18 can be alkylated at pyrazole position 1 with asuitable base, such as sodium hydride or potassium carbonate, in asuitable solvent, such as dimethylformamide, to yield the correspondingalkylated pyrazole derivatives 19. ##STR25##

General Synthetic Scheme V shows the preparation of a subset of thepyrazoles of the present invention where Q is an amide bridging radical.Ester 20 (prepared as set forth in Scheme VI) is hydrolyzed to thecorresponding acid, then converted to an acid halide, for example bytreatment with thionyl chloride. The acid halide is treated with asuitable alkali azide, such as sodium azide, and the resulting azideheated to effect the Curtius rearrangement to isocyanate 21. Isocyanate21 is hydrolyzed with aqueous acid to yield free amine 22, which can beoptionally alkylated to introduce the R⁶ substitution. Amine 22 is thenacylated with a suitable aromatic active ester or acid halide 23(preferably an aromatic active ester or acid chloride) to give pyrazoles24. ##STR26##

General Synthetic Scheme VI shows the preparation of a subset of thepyrazoles of Formula I where Q is an amide bridging radical. Optionallysubstituted 4-picoline 7 is deprotonated with a strong base, such aslithium hexamethyldisilazide, and acylated, for example, with oxalateester 25 to give pyruvate 26. Pyruvate 26 is reacted withdimethylformamide dimethyl acetal to give an intermediate vinyl aminethat is reacted with an optionally substituted hydrazine to givepyrazole ester 27. Pyrazole ester 27 is treated with aniline 28, underthe influence of heat, to give pyrazole 29 wherein R⁴ is preferablyhydrido. Alternatively, pyrazole ester 27 instead can be hydrolyzed toan intermediate acid which is then coupled to aniline 28 using aconventional amide coupling reaction (such as the coupling reaction ofpyrazole ester 27 with dicyclohexylcarbodiimide).

Pyruvate 26 also may be treated with a halogenating agent, such asN-chlorosuccinimide, to yield an intermediate haloketone that is thenreacted with thiosemicarbazide 30 to form pyrazole 31. Pyrazole 31 isaminated with aniline 28 using the procedure described above to produceaminopyrazole 32 wherein R¹ is hydrido, R⁴ is --NR¹⁵ R¹⁶, and R¹⁵ andR¹⁶ are, for example, hydrogen or alkyl, or together with the nitrogenatom to which they are attached form a 4 to 8 membered ring heterocyclylcomprising at least one heteroatom selected from oxygen, sulfur andnitrogen. The R¹ hydrido of aminopyrazole 32 can be replaced with othersubstituents as shown, for example, in Scheme IV. ##STR27##

General Synthetic Scheme VII shows the preparation of a subset of thepyrazoles of Formula I where Q is an ethanone bridging radical. Ester 33(prepared as set forth in Scheme VI) is treated with a Grignard reagent34 to produce pyrazole 35. Ester 33 may be suitably protected, employingprotecting groups known to those skilled in the art, to efficientlycarry out this transformation. ##STR28##

General Synthetic Scheme VIII shows the preparation of a subset of thepyrazoles of Formula I where Q is a cyclic bridging radical. Optionallysubstituted 4-picoline 7 is deprotonated with a strong base, such aslithium hexamethyldisilazide, and acylated with cyclic ester 36 to giveketone 37. Ketone 37 is reacted with dimethylformamide dimethyl acetalto give intermediate vinyl amine that is reacted with optionallysubstituted hydrazine to give pyrazole 38 wherein R⁴ is hydrido.

Alternatively, ketone 37 can be halogenated to form an intermediatehaloketone that can be reacted with optionally substitutedthiosemicarbazide 30 to give pyrazole 39 wherein R¹ is hydrido, R⁴ is--NR¹⁵ R¹⁶, and R¹⁵ and R¹⁶ are as defined for Scheme VI. ##STR29##

General Synthetic Scheme IX shows the preparation of a subset of thepyrazoles of Formula I where Q is a urea bridging radical. Isocyanate 40(prepared as set forth in Scheme V) is reacted with optionallysubstituted aniline 41 to give urea 42. ##STR30##

General Synthetic Scheme X shows the preparation of a subset of thepyrazoles of Formula I where Q is a sulfonamide bridging radical. Amine43 (prepared as set forth in Scheme V) is reacted with an optionallysubstituted aromatic sulfonyl halide 44, preferably sulfonyl chloride,to give pyrazole 45.

Within Formula I there is another subclass of compounds of interestrepresented by Formula III: ##STR31## Compounds of Formula III can beprepared in accordance with the chemistry set forth above (particularlyScheme V) and include the following compounds: ##STR32##

Within Formula I there is another subclass of compounds of interestrepresented by Formula IV: ##STR33##

Compounds of Formula IV can be prepared in accordance with the chemistryset forth above (particularly Scheme VI) and include the followingcompounds: ##STR34##

Within Formula I there is another subclass of compounds of interestrepresented by Formula V: ##STR35##

Compounds of Formula V can be prepared in accordance with the chemistryset forth above (particularly Scheme VII) and include the followingcompounds: ##STR36##

Within Formula I there is another subclass of compounds of interestrepresented by the Formula VI: ##STR37##

Compounds of Formula VI can be prepared in accordance with the chemistryset forth above (particularly Scheme VIII) and include the followingcompounds: ##STR38##

Within Formula I there is another subclass of compounds of interestrepresented by Formula VII: ##STR39##

Compounds of Formula VII can be prepared in accordance with thechemistry set forth above (particularly Scheme IX) and include thefollowing compounds: ##STR40##

Within Formula I there is another subclass of compounds of interestrepresented by Formula VIII ##STR41##

Compounds of Formula VIII can be prepared in accordance with thechemistry set forth above (particularly Scheme X) and include thefollowing compounds: ##STR42##

In the above compounds of Formula III, IV, V, VI, VII and VIII, thesubstituents R¹, R⁴, R⁵, R⁶ R⁷ R⁸, R⁹, R¹⁰ and R¹¹ are as previouslydefined for the compounds of Formula I, and substituent X1 encompassesthe optional substituents previously defined for the R² aryl group ofthe compounds of Formula I. In addition to the specific compoundsillustrated above, other specific compounds of Formula III, IV, V, VI,VII and VIII of particular interest include those wherein the R⁴hydrogen or methylpiperazinyl substituent is replaced withpropargylamino, substituted or unsubstituted piperidinyl, or substitutedor unsubstituted morpholinyl.

The following examples contain detailed descriptions of the methods ofpreparation of compounds of Formula I-VIII. These detailed descriptionsfall within the scope, and serve to exemplify, the above describedGeneral Synthetic Procedures which form part of the invention. Thesedetailed descriptions are presented for illustrative purposes only andare not intended as a restriction on the scope of the invention. Allparts are by weight and temperatures are in Degrees centigrade unlessotherwise indicated. All compounds showed NMR spectra consistent withtheir assigned structures. In some cases, the assigned structures wereconfirmed by nuclear Overhauser effect (NOE) experiments.

The following abbreviations are used:

MeOH--methanol

Pd/C--palladium on carbon

RT--room temperature

DTT--dithiotreitol

dH₂ O--distilled water

PBS--phosphate buffered saline

NaCl--sodium chloride

KCl--potassium chloride

Na₂ HPO₄ --sodium phosphate

KH₂ PO₄ --potassium phosphate

PMSF--phenylmethylsulfonyl fluoride

EDTA--ethylene diamine tetraacetic acid

HEPES--N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]

DMSO--dimethyl sulfoxide

SOCl₂ --thionyl chloride

NaN₃ --sodium azide

KOH--potassium hydroxide

Et--ethyl

LiHMDS--lithium hexamethyldisilazide

THF--tetrahydrofuran

DMF--dimethyl formamide

h--hour

min--minutes

EXAMPLE 1 ##STR43##4-[3-Methyl-5-(2-phenylethenyl)-1H-pyrazol-4-yl]pyridine

Step 1: Preparation of 6-phenyl-3-(4-pvridinvl)hexa-3,5-dien-2-one

Trans-cinnamaldehyde (1.84 ml, 14.8 mmol), five drops of piperidine andfive drops of acetic acid were added to a solution of 4-pyridyl acetone(2.0 g, 14.8 mmol) in toluene (25 ml). The mixture was heated to refluxfor 48 hours with a Dean-Stark trap to remove water. The mixture wasconcentrated to give a black oil. The oil was purified by silica gelchromatography (eluted with 1:1 hexane/ethyl acetate). The desiredfractions were collected, combined and concentrated to give6-phenyl-3-(4-pyridinyl)hexa-3,5-dien-2-one (628 mg). The ketone wasfurther purified by an additional column chromatography on silica gel(eluting with hexane-ethyl acetate-methylene chloride-methanol(1:1:2:0.12)) (380 mg, 27.3%): Anal. Calc'd for C₁₇ H₁₅ NO 0.4 H₂ O: C,79.91; H, 5.84; N, 5.48. Found: C, 79.72; H, 5.91; N, 5.25. MS (M+H⁺):250.

Step 2: Preparation of4-[3-methyl-5-(2-phenylethenyl)-1H-pyrazol-4-yl]pyridine

A solution of 6-phenyl-3-(4-pyridinyl)hexa-3,5-dien-2-one (step 1) (0.389, 1.52 mmol) and p-toluenesulfonyl hydrazide (0.28 g, 1.52 mmol) inacetic acid (3 ml) was heated to reflux for 48 hours. The mixture wasconcentrated and purified by silica gel chromatography (eluting with 1:1hexane/ethyl acetate). The desired fractions were collected, combinedand concentrated to give4-[3-methyl-5-(2-(phenylethenyl)-1H-pyrazol-4-yl]pyridine as anoff-white solid (26 mg, 6.5%): Anal. Calc'd for C₁₇ H₁₅ N₃ •0.75 H₂ O:C, 74.54; H, 6.03; N, 15.34. Found: C, 74.66; H, 5.69; 15.08. MS (M+H⁺):262 (100%).

EXAMPLE 2 ##STR44##4-[3-Methyl-5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine

4-(3-Methyl-5-(2-phenylethenyl)-1H-pyrazol-4-yl)pyridine (Example 1)(0.177 9, 0.67 mmol) was hydrogenated (5-30 psi) in MeOH in the presenceof a catalytic amount of 4% Pd/C at RT for 12 hours. The resultingmixture was filtered and the resulting solution was concentrated to give4-(3-methyl-5-phenylethyl-1H-pyrazol-4-yl)pyridine quantitatively: MS(M+H⁺) 264 (100%).

EXAMPLE 3 ##STR45## 4-[5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine

To a solution of lithium bis(trimethylsilyl)amide (100 mL, 1M, 100 mmol)in THF was added 4-picoline (9.73 mL, 100 mmol) at -10° C. The mixturewas warmed to 0° C. and held at this temperature for 1 hour. Ethylhydrocinnamate (8.82 mL, 50 mmol) was added slowly to the mixture at 0°C. The solution was stirred at 25° C. overnight. The mixture wasconcentrated, diluted with H₂ O (100 mL), neutralized with concentratedHCl to pH=8, and extracted with ethyl acetate (150 mL). The organiclayer was washed with H₂ O (100 mL×2), brine (50 mL), dried over MgSO₄,filtered and concentrated to give a yellow oil (11.2 g, 100% yield).

To the above oil (5.6 g, 25 mmol) dissolved in tetrahydrofuran (30 mL)was added N,N-dimethylformamide dimethyl acetal (13 mL, 98 mmol) and thesolution was stirred at room temperature for 48 hours. The solution wasconcentrated and dissolved in ethyl acetate (100 mL). The organic layerwas washed with H₂ O (50 mL×2), brine (50 mL), dried over MgSO₄,filtered and concentrated to give a yellow oil (2.6 g, 37% yield). Theoil (1.6 g) was purified by column chromatography on silica gel to givea pure product (0.36 g, 22%).

The above product (0.36 g, 1.3 mmol), hydrazine hydrate (0.4 mL, 12.7mmol) and methanol (2 mL) were mixed and stirred at 25° C. overnight.The mixture was concentrated and dissolved in ethyl acetate (20 mL). Theorganic layer was washed with H₂ O (10 mL×2), brine (10 mL), dried overMgSO₄, filtered and concentrated to give an oil. The oil (1.6 g) waspurified by column chromatography on silica gel to give4-[5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine (0.lg, 31.6% yield). mp:125-131 0C; mass spectrum (m/z): 250 (M+1). Anal. calc'd for C₁₆ H₁₅ N₃: C, 77.08; H, 6.28; N, 16.86. Found: C, 76.72; H, 6.28; N, none.

BIOLOGICAL EVALUATION

p38 Kinase Assay

Cloning of Human p38a:

The coding region of the human p38a CDNA was obtained byPCR-amplification from RNA isolated from the human monocyte cell lineTHP.1. First strand CDNA was synthesized from total RNA as follows: 2 μgof RNA was annealed to 100 ng of random hexamer primers in a 10 μlreaction by heating to 70° C. for 10 minutes followed by 2 minutes onice. cDNA was then synthesized by adding 1 μl of RNAsin (Promega,Madison Wis.), 2 μl of 50 mM dNTP's, 4 μl of 5×buffer, 2 μl of 100 mMDTT and 1 μl (200 U) of Superscript II™ AMV reverse transcriptase.Random primer, dNTP's and Superscript™ reagents were all purchased fromLife-Technologies, Gaithersburg, Mass. The reaction was incubated at 42°C. for 1 hour. Amplification of p38 cDNA was performed by aliquoting 5μl of the reverse transcriptase reaction into a 100 μl PCR reactioncontaining the following: 80 μl dH₂ O, 2 μl 50 mM dNTP's, 1 μl each offorward and reverse primers (50 pmol/μl), 10 μl of 10 ×buffer and 1 μlExpand™ polymerase (Boehringer Mannheim). The PCR primers incorporatedBam HI sites onto the 5' and 3' end of the amplified fragment, and werepurchased from Genosys. The sequences of the forward and reverse primerswere

5'-GATCGAGGATTCATCTCTCAGGAGAGGCCCA-3' and 5'GATCGAGGATTCTCAGGACTCCATCTCTTC-3' respectively. The PCR amplificationwas carried out in a DNA Thermal Cycler (Perkin Elmer) by repeating 30cycles of 94° C. for 1 minute, 60° C. for 1 minute and 68° C. for 2minutes. After amplification, excess primers and unincorporated dNTP'swere removed from the amplified fragment with a Wizard™ PCR prep(Promega) and digested with Bam HI (New England Biolabs). The Bam HIdigested fragment was ligated into BamHI digested pGEX 2T plasmid DNA(PharmaciaBiotech) using T-4 DNA ligase (New England Biolabs) asdescribed by T. Maniatis, Molecular Cloning: A Laboratory Manual, 2nded. (1989). The ligation reaction was transformed into chemicallycompetent E. coli DH10B cells purchased from Life-Technologies followingthe manufacturer's instructions. Plasmid DNA was isolated from theresulting bacterial colonies using a Promega Wizard™ miniprep kit.Plasmids containing the appropriate Bam HI fragment were sequenced in aDNA Thermal Cycler (Perkin Elmer) with Prism™ (Applied Biosystems Inc.).cDNA clones were identified that coded for both human p38a isoforms (Leeet al. Nature 372, 739). One of the clones which contained the cDNA forp38a-2 (CSBP-2) inserted in the cloning site of pGEX 2T, 3' of the GSTcoding region was designated pMON 35802. The sequence obtained for thisclone is an exact match of the cDNA clone reported by Lee et al. Thisexpression plasmid allows for the production of a GST-p38a fusionprotein.

Expression of Human p38a:

GST/p38a fusion protein was expressed from the plasmid PMON 35802 in E.coli,stain DH10B (Life Technologies, Gibco-BRL). Overnight cultures weregrown in Luria Broth (LB) containing 100 mg/ml ampicillin. The next day,500 ml of fresh LB was inoculated with 10 ml of overnight culture, andgrown in a 2 liter flask at 37° C. with constant shaking until theculture reached an absorbance of 0.8 at 600 nm. Expression of the fusionprotein was induced by addition of isopropyl b-D-thiogalactosidse (IPTG)to a final concentration of 0.05 mM. The cultures were shaken for threehours at room temperature, and the cells were harvested bycentrifugation. The cell pellets were stored frozen until proteinpurification.

Purification of p38 Kinase-α:

All chemicals were from Sigma Chemical Co. unless noted. Twenty grams ofE. coli cell pellet collected from five 1 L shake flask fermentationswas resuspended in a volume of PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 1.8 mM KH₂ PO₄, pH 7.3) up to 200 ml. The cell suspension wasadjusted to 5 mM DTT with 2 M DTT and then split equally into five 50 mlFalcon conical tubes. The cells were sonicated (Ultrasonics model W375)with a 1 cm probe for 3×1 minutes (pulsed) on ice. Lysed cell materialwas removed by centrifugation (12,000×g, 15 minutes) and the clarifiedsupernatant applied to glutathione-sepharose resin (Pharmacia).

Glutathione-Sepharose Affinity Chromatography:

Twelve ml of a 50% glutathione sepharose-PBS suspension was added to 200ml clarified supernatant and incubated batchwise for 30 minutes at roomtemperature. The resin was collected by centrifugation (600×g, 5 min)and washed with 2×150 ml PBS/1% Triton X-100, followed by 4×40 ml PBS.To cleave the p38 kinase from the GST-p38 fusion protein, theglutathione-sepharose resin was resuspended in 6 ml PBS containing 250units thrombin protease (Pharmacia, specific activity>7500 units/mg) andmixed gently for 4 hours at room temperature. The glutathione-sepharoseresin was removed by centrifugation (600×g, 5 min) and washed 2×6 mlwith PBS. The PBS wash fractions and digest supernatant containing p38kinase protein were pooled and adjusted to 0.3 mM PMSF.

Mono Q Anion Exchange Chromatography:

The thrombin-cleaved p38 kinase was further purified by FPLC-anionexchange chromatography. Thrombin-cleaved sample was diluted 2-fold withBuffer A (25 mM HEPES, pH 7.5, 25 mM beta-glycerophosphate, 2 mM DTT, 5%glycerol) and injected onto a Mono Q HR 10/10 (Pharmacia) anion exchangecolumn equilibrated width Buffer A. The column was eluted with a 160 ml0.1 M-0.6 M NaCl/Buffer A gradient (2 ml/minute flowrate). The p38kinase peak eluting at 200 mM NaCl was collected and concentrated to 3-4ml with a Filtron 10 concentrator (Filtron Corp.).

Sephacryl S100 Gel Filtration Chromatography:

The concentrated Mono Q- p38 kinase purified sample was purified by gelfiltration chromatography (Pharmacia HiPrep 26/60 Sephacryl S100 columnequilibrated with Buffer B (50 mM HEPES, pH 7.5, 50 mM NaCl, 2 mM DTT,5% glycerol)). Protein was eluted from the column with Buffer B at a 0.5ml/minute flowrate and protein was detected by absorbance at 280 nm.Fractions containing p38 kinase (detected by SDS-polyacrylamide gelelectrophoresis) were pooled and frozen at -80° C. Typical purifiedprotein yields from 5 L E. coli shake flasks fermentations were 35 mgp38 kinase.

In Vitro Assay

The ability of compounds to inhibit human p38 kinase alpha was evaluatedusing two in vitro assay methods. In the first method, activated humanp38 kinase alpha phosphorylates a biotinylated substrate, PHAS-I(phosphorylated heat and acid stable protein-insulin inducible), in thepresence of gamma ³² P-ATP (³² P-ATP). PHAS-I was biotinylated prior tothe assay and provides a means of capturing the substrate which isphosphorylated during the assay. p38 Kinase was activated by MKK6.Compounds were tested in 10 fold serial dilutions over the range of 100μM to 0.001 μM using 1% DMSO. Each concentration of inhibitor was testedin triplicate.

All reactions were carried out in 96 well polypropylene plates. Eachreaction well contained 25 mM HEPES pH 7.5, 10 mM magnesium acetate and50 μM unlabeled ATP. Activation of p38 was required to achievesufficient signal in the assay. Biotinylated PHAS-I was used at 1-2 μgper 50 μl reaction volume, with a final concentration of 1.5 μM.Activated human p38 kinase alpha was used at 1 μg per 50 μl reactionvolume representing a final concentration of 0.3 μM. Gamma ³² P-ATP wasused to follow the phosphorylation of PHAS-I. ³² P-ATP has a specificactivity of 3000 Ci/mmol and was used at 1.2 μCi per 50 μl reactionvolume. The reaction proceeded either for one hour or overnight at 30°C.

Following incubation, 20 Ml of reaction mixture was transferred to ahigh capacity streptavidin coated filter plate (SAM-streptavidin-matrix,Promega) prewetted with phosphate buffered saline. The transferredreaction mix was allowed to contact the streptavidin membrane of thePromega plate for 1-2 minutes. Following capture of biotinylated PHAS-Iwith ³² p incorporated, each well was washed to remove unincorporated ³²P-ATP three times with 2M NaCl, three washes of 2M NaCl with 1%phosphoric, three washes of distilled water and finally a single wash of95% ethanol. Filter plates were air dried and 20 μl of scintillant wasadded. The plates were sealed and counted. Results are shown in Table 4.

A second assay format was also employed that is based on p38 kinasealpha induced phosphorylation of EGFRP (epidermal growth factor receptorpeptide, a 21 mer) in the presence of ³³ P-ATP. Compounds were tested in10 fold serial dilutions over the range of 100 μM to 0.001 μM in 1%DMSO. Each concentration of inhibitor was tested in triplicate.Compounds were evaluated in 50 μl reaction volumes in the presence of 25mM Hepes pH 7.5, 10 mM magnesium acetate, 4% glycerol, 0.4% bovine serumalbumin, 0.4 mM DTT, 50 μM unlabeled ATP, 25 μg EGFRP (200 μM), and 0.05uCi gamma ³³ P-ATP. Reactions were initiated by addition of 0.09 μg ofactivated, purified human GST-p38 kinase alpha. Activation was carriedout using GST-MKK6 (5:1,p38:MKK6) for one hour at 30° C. in the presenceof 50 μM ATP. Following incubation for 60 minutes at room temperature,the reaction was stopped by addition of 150 μl of AG 1×8 resin in 900 mMsodium formate buffer, pH 3.0 (1 volume resin to 2 volumes buffer). Themixture was mixed three times with pipetting and the resin was allowedto settle. A total of 50 μl of clarified solution head volume wastransferred from the reaction wells to Microlite-2 plates. 150 μl ofMicroscint 40 was then added to each well of the Microlite plate, andthe plate was sealed, mixed, and counted.

                  TABLE I                                                         ______________________________________                                                      p38 kinase.sup.1                                                                        p38 kinase.sup.2                                      Example       IC50 μM                                                                              IC50 μM                                            ______________________________________                                        1             8.7       0.66                                                  2             1.0       2.8                                                   ______________________________________                                         .sup.1 p38α in vitro assay results based on PHASI assay procedure       .sup.2 p38α in vitro assay results based on EGFRP assay procedure  

TNF Cell Assays

Method of Isolation of Human Peripheral Blood Mononuclear Cells:

Human whole blood was collected in Vacutainer tubes containing EDTA asan anticoagulant. A blood sample (7 ml) was carefully layered over 5 mlPMN Cell Isolation Medium (Robbins Scientific) in a 15 ml round bottomcentrifuge tube. The sample was centrifuged at 450-500×g for 30-35minutes in a swing out rotor at room temperature. After centrifugation,the top band of cells were removed and washed 3 times with PBS w/ocalcium or magnesium. The cells were centrifuged at 400×g for 10 minutesat room temperature. The cells were resuspended in Macrophage Serum FreeMedium (Gibco BRL) at a concentration of 2 million cells/ml.

LPS Stimulation of Human PBMs:

PBM cells (0.1 ml, 2 million/ml) were co-incubated with 0.1 ml compound(10-0.41 μM, final concentration) for 1 hour in flat bottom 96 wellmicrotiter plates. Compounds were dissolved in DMSO initially anddiluted in TCM for a final concentration of 0.1% DMSO. LPS (Calbiochem,20 ng/ml, final concentration) was then added at a volume of 0.010 ml.Cultures were incubated overnight at 37° C. Supernatants were thenremoved and tested by ELISA for TNF-a and IL1-b. Viability was analyzedusing MTS. After 0.1 ml supernatant was collected, 0.020 ml MTS wasadded to remaining 0.1 ml cells. The cells were incubated at 37° C. for2-4 hours, then the O.D. was measured at 490-650 nM.

Maintenance and Differentiation of the U937 Human Histiocytic LvmphomaCell Line:

U937 cells (ATCC) were propagated in RPMI 1640 containing 10% fetalbovine serum, 100 IU/ml penicillin, 100 μg/ml streptomycin, and 2 mMglutamine (Gibco). Fifty million cells in 100 ml media were induced toterminal monocytic differentiation by 24 hour incubation with 20 ng/mlphorbol 12-myristate 13-acetate (Sigma). The cells were washed bycentrifugation (200×g for 5 min) and resuspended in 100 ml fresh medium.After 24-48 hours, the cells were harvested, centrifuged, andresuspended in culture medium at 2 million cells/ml.

LPS Stimulation of TNF production by U937 Cells:

U937 cells (0.1 ml, 2 million/ml) were incubated with 0.1 ml compound(0.004-50 μM, final concentration) for 1 hour in 96 well microtiterplates. Compounds were prepared as 10 mM stock solutions in DMSO anddiluted in culture medium to yield a final DMSO concentration of 0.1% inthe cell assay. LPS (E coli, 100 ng/ml final concentration) was thenadded at a volume of 0.02 ml. After 4 hour incubation at 37° C., theamount of TNF-α released in the culture medium was quantitated by ELISA.Inhibitory potency is expressed as IC50 (μM). Results are shown in TableII.

                  TABLE II                                                        ______________________________________                                                      TNF Cell                                                               Example                                                                              IC50 μM                                                      ______________________________________                                               1      0.6                                                                    2      2.3                                                             ______________________________________                                    

Rat Assay

The efficacy of the novel compounds in blocking the production of TNFalso was evaluated using a model based on rats challenged with LPS. MaleHarlen Lewis rats [Sprague Dawley Co.] were used in this model. Each ratweighed approximately 300 g and was fasted overnight prior to testing.Compound administration was typically by oral gavage (althoughintraperitoneal, subcutaneous and intravenous administration were alsoused in a few instances) 1 to 24 hours prior to the LPS challenge. Ratswere administered 30 μg/kg LPS [salmonella typhosa, Sigma Co.]intravenously via the tail vein.

Blood was collected via heart puncture 1 hour after the LPS challenge.Serum samples were stored at -20° C. until quantitative analysis ofTNF-α by Enzyme Linked-Immuno-Sorbent Assay ("ELISA") [Biosource].Additional details of the assay are set forth in Perretti, M., et al.,Br. J. Pharmacol. (1993), 110, 868-874, which is incorporated byreference in this application.

Mouse Assay

Mouse Model Of LPS-Induced TNF Alpha Production:

TNF alpha was induced in 10-12 week old BALB/c female mice by tail veininjection with 100 ng lipopolysaccharide (from S. Typhosa) in 0.2 mlsaline. One hour later mice were bled from the retroorbital sinus andTNF concentrations in serum from clotted blood were quantified by ELISA.Typically, peak levels of serum TNF ranged from 2-6 ng/ml one hour afterLPS injection.

The compounds tested were administered to fasted mice by oral gavage asa suspension in 0.2 ml of 0.5% methylcellulose and 0.025% Tween 20 inwater at 1 hour or 6 hours prior to LPS injection. The 1 hour protocolallowed evaluation of compound potency at Cmax plasma levels whereas the6 hour protocol allowed estimation of compound duration of action.Efficacy was determined at each time point as percent inhibition ofserum TNF levels relative to LPS injected mice that received vehicleonly.

Induction And Assessment Of Collagen-Induced Arthritis In Mice:

Arthritis was induced in mice according to the procedure set forth in J.M. Stuart, Collagen Autoimmune Arthritis, Annual Rev. Immunol. 2:199(1984), which is incorporated herein by reference. Specifically,arthritis was induced in 8-12 week old DBA/1 male mice by injection of50 μg of chick type II collagen (CII) (provided by Dr. Marie Griffiths,Univ. of Utah, Salt Lake City, Utah) in complete Freund's adjuvant(Sigma) on day 0 at the base of the tail. Injection volume was 100 μl.Animals were boosted on day 21 with 50 μg of CII in incomplete Freund'sadjuvant (100 μl volume). Animals were evaluated several times each weekfor signs of arthritis. Any animal with paw redness or swelling wascounted as arthritic. Scoring of arthritic paws was conducted inaccordance with the procedure set forth in Wooley et al., GeneticControl of Type II Collagen Induced Arthritis in Mice: FactorsInfluencing Disease Suspectibility and Evidence for Multiple MHCAssociated Gene Control., Trans. Proc., 15:180 (1983). Scoring ofseverity was carried out using a score of 1-3 for each paw (maximalscore of 12/mouse). Animals displaying any redness or swelling of digitsor the paw were scored as 1. Gross swelling of the whole paw ordeformity was scored as 2. Ankylosis of joints was scored as 3. Animalswere evaluated for 8 weeks. 8-10 animals per group were used.

Preparation and Administration of Compounds:

The compounds tested on mice having collagen-induced arthritis wereprepared as a suspension in 0.5% methylcelluose (Sigma, St. Louis, Mo.),0.025% Tween 20 (Sigma). The compound suspensions were administered byoral gavage in a volume of 0.1 ml b.i.d. Administration began on day 20post collagen injection and continued daily until final evaluation onday 56. Scoring of arthritic paws was conducted as set forth above.

Also embraced within this invention is a class of pharmaceuticalcompositions comprising the active compounds of this invention inassociation with one or more non-toxic, pharmaceutically-acceptablecarriers and/or diluents and/or adjuvants (collectively referred toherein as "carrier" materials) and, if desired, other activeingredients. The active compounds of the present invention may beadministered by any suitable route, preferably in the form of apharmaceutical composition adapted to such a route, and in a doseeffective for the treatment intended. The active compounds andcomposition may, for example, be administered orally, intravascularly(IV), intraperitoneally, subcutaneously, intramuscularly (IM) ortopically. For oral administration, the pharmaceutical composition maybe in the form of, for example, a tablet, hard or soft capsule,lozenges, dispensable powders, suspension or liquid. The pharmaceuticalcomposition is preferably made in the form of a dosage unit containing aparticular amount of the active ingredient. Examples of such dosageunits are tablets or capsules. The active ingredient may also beadministered by injection (IV, IM, subcutaneous or jet) as a compositionwherein, for example, saline, dextrose, or water may be used as asuitable carrier. The pH of the composition may be adjusted, ifnecessary, with suitable acid, base, or buffer. Suitable bulking,dispersing, wetting or suspending agents, including mannitol and PEG400, may also be included in the composition. A suitable parenteralcomposition can also include a compound formulated as a sterile solidsubstance, including lyophilized powder, in injection vials. Aqueoussolution can be added to dissolve the compound prior to injection. Theamount of therapeutically active compounds that are administered and thedosage regimen for treating a disease condition with the compoundsand/or compositions of this invention depends on a variety of factors,including the age, weight, sex and medical condition of the subject, theseverity of the inflammation or inflammation related disorder, the routeand frequency of administration, and the particular compound employed,and thus may vary widely. The pharmaceutical compositions may containactive ingredients in the range of about 0.1 to 1000 mg, preferably inthe range of about 7.0 to 350 mg. A daily dose of about 0.01 to 100mg/kg body weight, preferably between about 0.1 and about 50 mg/kg bodyweight and most preferably between about 0.5 to 30 mg/kg body weight,may be appropriate. The daily dose can be administered in one to fourdoses per day. In the case of skin conditions, it may be preferable toapply a topical preparation of compounds of this invention to theaffected area two to four times a day. For disorders of the eye or otherexternal tissues, e.g., mouth and skin, the formulations are preferablyapplied as a topical gel, spray, ointment or cream, or as a suppository,containing the active ingredients in a total amount of, for example,0.075 to 30% w/w, preferably 0.2 to 20% w/w and most preferably 0.4 to15% w/w. When formulated in an ointment, the active ingredients may beemployed with either paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream withan oil-in-water cream base. If desired, the aqueous phase of the creambase may include, for example at least 30% w/w of a polyhydric alcoholsuch as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol,polyethylene glycol and mixtures thereof. The topical formulation maydesirably include a compound which enhances absorption or penetration ofthe active ingredient through the skin or other affected areas. Examplesof such dermal penetration enhancers include dimethylsulfoxide andrelated analogs. The compounds of this invention can also beadministered by a transdermal device. Preferably topical administrationwill be accomplished using a patch either of the reservoir and porousmembrane type or of a solid matrix variety. In either case, the activeagent is delivered continuously from the reservoir or microcapsulesthrough a membrane into the active agent permeable adhesive, which is incontact with the skin or mucosa of the recipient. If the active agent isabsorbed through the skin, a controlled and predetermined flow of theactive agent is administered to the recipient. In the case ofmicrocapsules, the encapsulating agent may also function as themembrane. The transdermal patch may include the compound in a suitablesolvent system with an adhesive system, such as an acrylic emulsion, anda polyester patch. The oily phase of the emulsions of this invention maybe constituted from known ingredients in a known manner. While the phasemay comprise merely an emulsifier, it may comprise a mixture of at leastone emulsifier with a fat or an oil or with both a fat and an oil.Preferably, a hydrophilic emulsifier is included together with alipophilic emulsifier which acts as a stabilizer. It is also preferredto include both an oil and a fat. Together, the emulsifier(s) with orwithout stabilizer(s) make-up the so-called emulsifying wax, and the waxtogether with the oil and fat make up the so-called emulsifying ointmentbase which forms the oily dispersed phase of the cream formulations.Emulsifiers and emulsion stabilizers suitable for use in the formulationof the present invention include Tween 60, Span 80, cetostearyl alcohol,myristyl alcohol, glycerol monostearate, and sodium lauryl sulfate,among others. The choice of suitable oils or fats for the formulation isbased on achieving the desired cosmetic properties, since the solubilityof the active compound in most oils likely to be used in pharmaceuticalemulsion formulations is very low. Thus, the cream should preferably bea non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as di-isoadipate,isocetyl stearate, propylene glycol diester of coconut fatty acids,isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters may be used.These may be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils can be used.Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredients are dissolved or suspended insuitable carrier, especially an aqueous solvent for the activeingredients. The anti-inflammatory active ingredients are preferablypresent in such formulations in a concentration of 0.5 to 20%,advantageously 0.5 to 10% and particularly about 1.5% w/w. Fortherapeutic purposes, the active compounds of this combination inventionare ordinarily combined with one or more adjuvants appropriate to theindicated route of administration. If administered per os, the compoundsmay be admixed with lactose, sucrose, starch powder, cellulose esters ofalkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesiumstearate, magnesium oxide, sodium and calcium salts of phosphoric andsulfuric acids, gelatin, acacia gum, sodium alginate,polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted orencapsulated for convenient administration. Such capsules or tablets maycontain a controlled-release formulation as may be provided in adispersion of active compound in hydroxypropylmethyl cellulose.Formulations for parenteral administration may be in the form of aqueousor non-aqueous isotonic sterile injection solutions or suspensions.These solutions and suspensions may be prepared from sterile powders orgranules having one or more of the carriers or diluents mentioned foruse in the formulations for oral administration. The compounds may bedissolved in water, polyethylene glycol, propylene glycol, ethanol, cornoil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodiumchloride, and/or various buffers. Other adjuvants and modes ofadministration are well and widely known in the pharmaceutical art.

All patent documents listed herein are incorporated by reference.

Although this invention has been described with respect to specificembodiments, the details of these embodiments are not to be construed aslimitations.

What we claim is:
 1. A compound of Formula IA ##STR46## wherein R¹ isselected from hydrido, lower alkyl, lower alkynyl, lowercycloalkylalkylene, lower haloalkyl, lower hydroxyalkyl, loweralkoxyalkyl, lower thioalkyl, lower alkylthioalkylene, loweralkylaminoalkylene, and lower heterocyclylalkylene; andQ is selectedfrom lower alkylene, lower alkenylene, ##STR47## represents a four toeight membered ring heterocyclylidenyl comprising one or moreheteroatoms selected from oxygen, sulfur and nitrogen; andwherein n isan integer from 1 to 7; and R² is aryl optionally substituted with oneor more radicals independently selected from halo, lower alkyl, loweralkoxy, lower aryloxy, lower aralkoxy, amino, hydroxyl, nitro, cyano,lower haloalkyl, lower alkylamino, and lower alkynylamino; and R³ ispyridyl optionally substituted with one or more radicals independentlyselected from halo, lower alkyl, cyano, phenethyl, benzyl, benzyloxy,benzylthio, benzylamino, phenethylamino, aminocarbonyl, loweralkylcarbonylamino, hydroxyl, amino, lower alkylamino, lower aminoalkyl,and phenylamino; and R⁴ is selected from hydrido, lower alkyl, phenyl,lower haloalkyl, 5-10 membered heterocyclyl, lower alkylamino, loweralkynylamino, phenylamino, lower cycloalkyl, lower alkenyl, lowercycloalkenyl, lower alkoxy, lower heterocyclylaminoalkyl, and lowerheterocyclylalkylamino; wherein the phenyl, 5-10 membered heteroaryl,lower cycloalkyl and lower cycloalkenyl groups are optionallysubstituted with one or more radicals independently selected from halo,lower alkyl, lower alkenyl, lower alkynyl, lower alkoxy, phenoxy, loweraralkoxy, lower haloalkyl, amino, hydroxyl, cyano and lower alkylamino;and wherein the amino radicals of lower heterocyclyl alkylamino andlower heterocyclylaminoalkyl are optionally substituted with one or morelower alkyl; and R⁶ is selected from hydrido, lower alkyl, loweralkenyl, and lower alkynyl, and R⁷ and R⁸ are independently selectedfrom hydrido, lower lower alkyl, lower alkenyl, and lower alkynyl, ortogether form a carbocyclic or heterocyclic ring having three to eightmembers; and R⁹ is selected from hydrido, lower alkyl, lower alkenyl,and lower alkynyl; and R¹⁰ is selected from hydrido, lower alkyl, loweralkenyl, and lower alkynyl; and R¹¹ is selected from hydrido, loweralkyl, lower alkenyl, and lower alkynyl; and R¹² is selected fromhydrido and lower alkyl; or a pharmaceutically acceptable salt thereof.2. Compounds of claim 1 whereinR¹ is selected from hydrido, methyl,ethyl, propyl, isopropyl, tert-butyl, isobutyl, ethynyl, propargyl,fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl,dichloromethyl, trichloroethyl, pentafluoroethyl, heptafluoropropyl,difluorochloromethyl, dichlorofluoromethyl, difluoroethyl,difluoropropyl, dichloroethyl, dichloropropyl, morpholinomethyl,pyrrolidinylmethyl, piperazinylmethyl, piperidinylmethyl, pyridylmethyl,thienylmethyl, methoxymethyl, ethoxymethyl, methylaminomethyl,cyclohexylmethyl, hydroxymethyl, hydroxylethyl, thiomethyl, andmethylthiomethyl; and Q is selected from methylene, ethylene, propylene,ethenylene, propenylenyl ##STR48## represents a four to eight memberedring heterocyclylidenyl comprising one or more heteroatoms selected fromoxygen, sulfur and nitrogen; andwherein n is an integer from 1 to 7; andR² is phenyl optionally substituted with one or more radicalsindependently selected from fluoro, chloro, bromo, methyl, ethyl,isopropyl, tert-butyl, isobutyl, methoxy, ethoxy, phenoxy, benzyloxy,trifluoromethyl, fluoromethyl, difluoromethyl, amino, cyano, nitro,dimethylamino, ethynylamino, propargylamino, and hydroxyl; and R³ ispyridyl optionally substituted with one or more radicals independentlyselected from fluoro, chloro, bromo, methyl, ethyl, isopropyl, cyano,aminocarbonyl, methylcarbonylamino, hydroxy, benzyl, phenethyl,methylamino, ethylamino, dimethylamino, diethylamino, aminomethyl,aminoethyl, N-methyl-N-phenylamino, phenylamino, diphenylamino,benzylamino, phenethylamino, and amino; and R⁴ is selected from hydrido,methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, isobutyl, phenyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylenyl,cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, pyridyl,thienyl, isothiazolyl, isoxazolyl, thiazolyl, oxazolyl, pyrimidinyl,quinolyl, isoquinolinyl, imidazolyl, benzimidazolyl, furyl, benzofuryl,methoxy, ethoxy, trifluoromethyl, fluoromethyl, methylamino,ethynylamino, propargylamino, piperidinyl, piperazinyl,piperadinylmethyl, piperadinylmethylamino, piperadinylaminomethyl,piperazinylmethyl, piperazinylmethylamino, piperazinylaminomethyl;wherein the phenyl, piperadinyl and piperazinyl groups are optionallysubstituted with one or more radicals independently selected fromfluoro, chloro, bromo, methyl, ethyl, isopropyl, tert-butyl, isobutyl,propargyl, methoxy, ethoxy, phenoxy, benzyloxy, trifluoromethyl,fluoromethyl, difluoromethyl, amino, hydroxyl, cyano and dimethylamino;and wherein the amino radicals of piperadinylmethylamino,piperadinylaminomethyl, piperazinylmethylamino, andpiperazinylaminomethyl are optionally substituted with one or moremethyl; and R⁶ is selected from hydrido, methyl, ethyl, propyl,isopropyl, tert-butyl, and isobutyl; and R⁷ and R⁸ are independentlyselected from hydrido, methyl, ethyl, propyl, isopropyl, tert-butyl, andisobutyl, or together form a carbocyclic or heterocyclic ring havingthree to eight members; and R⁹ is selected from hydrido, methyl, ethyl,propyl, isopropyl, tert-butyl, and isobutyl; and R¹⁰ is selected fromhydrido, methyl, ethyl, propyl, isopropyl, tert-butyl, and isobutyl; andR¹¹ is selected from hydrido, methyl, ethyl, propyl, isopropyl,tert-butyl, and isobutyl; and R¹² is selected from hydrido, methyl,ethyl, propyl, isopropyl, tert-butyl, and isobutyl; or apharmaceutically-acceptable salt thereof.
 3. Compound of claim 2whereinR¹ is hydrido or methyl; and Q is selected from methylene,ethylene, ethenylene, ##STR49## represents a four to eight membered ringheterocyclylidenyl comprising one or more heteroatoms selected fromoxygen, sulfur and nitrogen; andwherein n is an integer from 1 to 3; andR² is phenyl optionally substituted with one or more radicalsindependently selected from fluoro, chloro, bromo, methyl, ethyl,isopropyl, tert-butyl, isobutyl, methoxy, ethoxy, phenoxy, benzyloxy,trifluoromethyl, fluoromethyl, difluoromethyl, amino, cyano, nitro,dimethylamino, and hydroxyl; and R³ is pyridyl optionally substitutedwith one or more radicals independently selected from fluoro, chloro,bromo, methyl, cyano, benzyl, phenethyl, aminocarbonyl, hydroxyl,dimethylamino, benzylamino, phenethylamino, aminomethyl and amino; andR⁴ is selected from hydrido, methyl, ethyl, propyl, propargylamino, andphenyl optionally substituted with one or more radicals independentlyselected from fluoro, chloro, bromo, methyl, ethyl, isopropyl, methoxy,ethoxy, phenoxy, benzyloxy, trifluoromethyl, dimethylamino, ethynylaminoand propargylamino; and R⁶ is selected from hydrido and methyl; and R⁷and R⁸ are independently selected from hydrido and methyl; and R⁹ isselected from hydrido and methyl; and R¹⁰ is selected from hydrido andmethyl; and R¹¹ is selected from hydrido and methyl; and R¹² is selectedfrom hydrido and methyl; or a pharmaceutically-acceptable salt.
 4. Acompound, or its tautomer or a pharmaceutically acceptable salt thereof,selected from the group consistingof4-[3-methyl-5-(2-phenylethenyl)-1H-pyrazol-4-yl]pyridine;4-[3-methyl-5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine;4-[3-methyl-5-[2-(3-fluorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;4-[3-[2-(3-fluorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;4-[1-methyl-3-[2-(3-fluorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;4-[3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;4-[l-methyl-3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;4-[3-[2-(3-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;3-methyl-4-[l-methyl-3-[2-(3,4-dichlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-[2-(4-chlorophenyl)ethyl]-1H-pyrazol-4-yl]pyridine;4-[3-methyl-5-[2-(3-fluorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;4-[3-[2-(3-fluorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;4-[1-methyl-3-[2-(3-fluorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;4-[3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;4-[1-methyl-3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine4-[3-[2-(3-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-[2-(3,4-dichlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-[2-(4-chlorophenyl)ethenyl]-1H-pyrazol-4-yl]pyridine;4-[3-methyl-5-(3-fluorobenzyl)-1H-pyrazol-4-yl]pyridine;4-[3-(3-fluorobenzyl)-1H-pyrazol-4-yl]pyridine;4-[1-methyl-3-(3-fluorobenzyl)-1H-pyrazol-4-yl]pyridine;4-[3-benzyl-1H-pyrazol-4-yl]pyridine;4-[3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;4-[1-methyl-3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;4-[3-(3-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-(3,4-dichlorobenzyl)-1H-pyrazol-4-yl]pyridine;3-methyl-4-[1-methyl-3-(4-chlorobenzyl)-1H-pyrazol-4-yl]pyridine;(3-fluorophenyl)[3-methyl-4-(4-pyridinyl)-1H-pyrazol-5-yl]methanone;(3-fluorophenyl)[4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;(3-fluorophenyl)[1-methyl-4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;phenyl[1-methyl-4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;(4-chlorophenyl)[4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;(4-chlorophenyl)[1-methyl-4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;(4-chlorophenyl)[1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazol-3-yl]methanone;(3-chlorophenyl)[4-(4-pyridinyl)-1H-pyrazol-3-yl]methanone;(3,4-dichlorophenyl)[1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazol-3-yl]methanone;(4-chlorophenyl)[1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazol-3-yl]methanone;α-(3-fluorophenyl)-3-methyl-4-(4-pyridinyl)-1H-pyrazole-5-methanol;α-(3-fluorophenyl)-4-(4-pyridinyl)-1H-pyrazole-3-methanol;α-(3-fluorophenyl)-1-methyl-4-(4-pyridinyl)-1H-pyrazole-3-methanol;α-phenyl-1-methyl-4-(4-pyridinyl)-1H-pyrazole-3-methanol;α-(4-chlorophenyl)-4-(4-pyridinyl)-1H-pyrazole-3-methanol;α-(4-chlorophenyl)-1-methyl-4-(4-pyridinyl)-1H-pyrazole-3-methanol;α-(4-chlorophenyl)-1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazole-3-methanol;α-(3-chlorophenyl)-4-(4-pyridinyl)-1H-pyrazole-3-methanol;α-(3,4-dichlorophenyl)-1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazole-3-methanol;α-(4-chlorophenyl)-1-methyl-4-(3-methyl-4-pyridinyl)-1H-pyrazole-3-methanol;and 4-[5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine.
 5. A compound, or itstautomer or a pharmaceutically acceptable salt thereof, selected fromthe group consisting of ##STR50##
 6. A compound of Formula II whereinR¹is selected from hydrido and lower alkyl; and Q is selected from loweralkeylene, lower alkenylene, ##STR51## represents a four to eightmembered ring heterocyclylidenyl comprising one or more heteroatomsselected from oxygen, sulfur and nitrogen; and wherein n is an integerfrom 1 to 4; and R² is phenyl optionally substituted with one or moreradicals independently selected from halo and lower alkyl; and R⁴, R⁵,R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, and R¹² are independently selected fromhydrido and lower alkyl; or a pharmaceutically-acceptable salt ortautomer thereof.
 7. Compound of claim 6 whereinR¹ is selected fromhydrido and methyl; and wherein Q is selected from methylene, ethylene,ethenylene, ##STR52## R² is phenyl optionally substituted with one ormore radicals independently selected from fluoro, chloro and bromo; andR⁴ is selected from hydrido, methyl and ethyl; and R⁵ is selected fromhydrido and methyl; or a pharmaceutically-acceptable salt or tautomerthereof.
 8. A compound of claim 6, or its tautomer or a pharmaceuticallyacceptable salt thereof, selected from the group consistingof4-[3-methyl-5-(2-phenylethenyl)-1H-pyrazol-4-yl]pyridine;4-[3-methyl-5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine; and4-[5-(2-phenylethyl)-1H-pyrazol-4-yl]pyridine.
 9. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 1; or apharmaceutically-acceptable salt thereof.
 10. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 2; or apharmaceutically-acceptable salt thereof.
 11. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 3; or apharmaceutically-acceptable salt thereof.
 12. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 4; or apharmaceutically-acceptable salt thereof.
 13. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 5; or apharmaceutically-acceptable salt thereof.
 14. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 6; or apharmaceutically-acceptable salt thereof.
 15. A pharmaceuticalcomposition comprising a therapeutically-effective amount of a compound,said compound selected from a family of compounds of claim 7; or apharmaceutically-acceptable salt or tautomer thereof.
 16. A method oftreating a TNF mediated disorder, said method comprising treating thesubject having or susceptible to such disorder with atherapeutically-effective amount of a compound of claim 1; or apharmaceutically-acceptable salt or tautomer thereof.
 17. A method oftreating a p38 kinase mediated disorder, said method comprising treatingthe subject having or susceptible to such disorder with atherapeutically-effective amount of a compound of claim 1; or apharmaceutically-acceptable salt or tautomer thereof.
 18. A method oftreating inflammation, said method comprising treating the subjecthaving or susceptible to inflammation with a therapeutically-effectiveamount of a compound of claim 1; or a pharmaceutically-acceptable saltor tautomer thereof.
 19. A method of treating arthritis, said methodcomprising treating the subject having or susceptible to arthritis witha therapeutically-effective amount of a compound of claim 1; or apharmaceutically-acceptable salt or tautomer thereof.
 20. The method ofclaim 16 wherein the p38 TNF mediated disorder is selected from thegroup of disorders consisting of bone resorption, graft vs. hostreaction, atherosclerosis, arthritis, osteoarthritis, rheumatoidarthritis, gout, psoriasis, topical inflammatory disease state, adultrespiratory distress syndrome, asthma, chronic pulmonary inflammatorydisease, cardiac reperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Crohn's disease, ulcerative colitis, inflammatorybowel disease and cachexia.
 21. The method of claim 16 wherein the TNFmediated disorder is inflammation.
 22. The method of claim 16 whereinthe TNF mediated disorder is arthritis.
 23. The method of claim 16wherein the TNF mediated disorder is asthma.
 24. The method of claim 17wherein the disorder is a p38α kinase mediated disorder.
 25. The methodof claim 17 wherein the p38 kinase mediated disorder is selected fromthe group of diseases consisting of bone resorption, graft vs. hostreaction, atherosclerosis, arthritis, osteoarthritis, rheumatoidarthritis, gout, psoriasis, topical inflammatory disease state, adultrespiratory distress syndrome, asthma, chronic pulmonary inflammatorydisease, cardiac reperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Crohn's disease, ulcerative colitis, inflammatorybowel disease and cachexia.
 26. The method of claim 17 wherein the p38kinase mediated disorder is inflammation.
 27. The method of claim 17wherein the p38 kinase mediated disorder is arthritis.
 28. The method ofclaim 17 wherein the p38 kinase mediated disorder is asthma.
 29. Amethod of treating a TNF medicated disorder, said method comprisingtreating the subject having or susceptible to such disorder with atherapeutically-effective amount of a compound of claim 6; or apharmaceutically-acceptable salt or tautomer thereof.
 30. A method oftreating a p38 kinase mediated disorder said method comprising treatingthe subject having or susceptible to such disorder with atherapeutically-effective amount of a compound of claim 6; or apharmaceutically-acceptable salt of tautomer thereof.
 31. A method oftreating inflammation, said method comprising treating the subjecthaving or susceptible to inflammation with a therapeutically-effectiveamount of a compound of claim 6; or a pharmaceutically-acceptable saltor tautomer thereof.
 32. A method of treating arthritis, said methodcomprising treating the subject having or susceptible to arthritis witha therapeutically-effective amount of a compound of claim 6; or apharmaceutically acceptable salt or tautomer thereof.
 33. The method ofclaim 29 wherein the p38 TNF mediated disorder is selected from thegroup of disorders consisting of bone resorption, graft vs. hostreaction, atherosclerosis, arthritis, osteoarthritis, rheumatoidarthritis, gout, psoriasis, topical inflammatory disease state, adultrespiratory distress syndrome, asthma, chronic pulmonary inflammatorydisease, cardiac reperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Crohn's disease, ulcerative colitis, inflammatorybowel disease and cachexia.
 34. The method of claim 29 wherein the TNFmediated disorder is inflammation.
 35. The method of claim 29 whereinthe TNF mediated disorder is arthritis.
 36. The method of claim 29wherein the TNF mediated disorder is asthma.
 37. The method of claim 30wherein the disorder is a p38α kinase mediated disorder.
 38. The methodof claim 30 wherein the p38 kinase mediated disorder is selected fromthe group of diseases consisting of bone resorption, graft vs. hostreaction, atherosclerosis, arthritis, osteoarthritic, rheumatoidarthritis, gout, psoriasis, topical inflammatory disease stato, adultrespiratory distress syndrome, asthma, chronic pulmonary inflammatorydisease, cardiac reperfusion injury, renal reperfusion injury, thrombus,glomerulonephritis, Crohn's disease, ulcerative colitis, inflamatorybowel disease and cachexia.
 39. The method of claim 30 wherein the p38kinase mediated disorder is inflammation.
 40. The method of claim 30wherein the p38 kinase mediated disorder is arthritis.
 41. The method ofclaim 30 wherein the p38 kinase mediated disorder is asthma.